2.11 The secret origin of the new Marvel Universe.The Celestials debuted in The Eternals #1 (July 1976) and were created by writer and artist Jack Kirby. They reappeared as regular guest stars in three subsequent limited series sequels: The Eternals vol. The characters have also been featured in other titles, including the "Celestial Saga" storyline in Thor Annual #7 (1978), Thor #283–300 (May 1979–Oct. The first detailed account of the Celestials' origin was finally presented in The Ultimates 2 #6 (2017). The origin of the Celestials had long been unknown, with many species across the mainstream Marvel Universe having only legends about their beginning, none of which have been validated, until the so-called Eternity War, during which major revelations about the origin and nature of the Celestials were revealed by the mysterious cosmic entity called the Queen of Nevers.Īt the beginning of creation itself, countless billions of years ago, before the current Cosmic Order, creation was composed of a single and sentient universe, whose omnipotent intelligence was referred to as the First Firmament. For countless ages, the First Firmament was the sole being in creation, until its loneliness became unbearable. It decided to create the first life in Creation to give it companions as well as servants-an act that it would later come to regret. These servants were cosmic beings of a lesser order of power and were of two kinds: black and multicolored humanoid servants. The black servants dutifully obeyed and worshiped their creator. They even created their own servants and sought to preserve the simple order their creator had made complete and unchanging for all time. The First Firmament named these loyal beings Aspirants and was very pleased by their goals and desire to maintain the status quo of its reign. However, the multicolored ones had completely different values and desires from the Aspirants. ![]() Considered "rebels" by the First Firmament, they wanted a dynamic, diverse and continually evolving Reality where beings lived, learned, reproduced, aged and died in order to slowly improve themselves through evolution. The rebels wanted this with the ultimate long-term goal of producing superior cosmic beings with the power to create universes of their own and for the universe to evolve with them as they advanced towards that state. These were the beings whom one day would be called by lesser life forms, "The Celestials". The two opposing factions of the First Firmament's children could not peacefully co-exist and the ensuing war nearly destroyed the first universe. At some point during the war, the Aspirants created a now-lost hyper weapon called the Godkiller, a space-borne 25,000 foot (7,600 m) tall humanoid robot that dwarfed even the Celestials themselves. It was powered by a cosmic artifact later called the Heart of The Voldi (named after the species which would adopt it) and operated by genetically engineered pilots.
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![]() There's new modes as well, including a Gatecrasher mode where you'll smash through as many barriers as you can which each add 2 seconds to your time limit, and the racer with the highest limit at the end wins. You can change the difficulty each race for for cash and experience.even on Casual difficulty this a "don't make 1 mistake" kind of game, but it's realism stops short of boring stuff like having to pay for repairs. Same with the accelerator, and you really notice a difference in how that punishes you on the different surfaces, from the narrow, gravelly tracks of Croatia, to sandy Baha, and even the streets of Shibuya. Driving also seems a little less forgiving on the turns, and you'll have to ween yourself off that tendency to push the joystick all the way to the side, and practice slight movements. Yeah, not to mention the cars themselves. It was hard to believe they could improve on the first DiRT in the looks department, but they've done it. But rather than take a different direction, the latest in the franchise is a tribute to the racing legend, and Colin McRae DiRT 2 pays plenty of homage.Įvery now and then a game comes along and suprises you with how good it looks, and DiRT 2 will render scenery that makes you proud of your hardware, be it 360, PS3, or PC. Shortly after, Colin McRae himself died tragically with his son in a helicopter accident. A positive value will tilt the camera up.Several years ago, Codemasters released a masterpiece of rally racing realism, in the form of Colin McRae: DiRT.Compared to the position values, the tilt value works on a different scale: a modification of 1 will yield a barely noticeable change. Head Tilt Change: affects the vertical angle of the camera.A positive value will move the point of view closer to the dashboard. A positive value will move the point of view upward. A positive value will move the point of view to the left. It is to be noted that the point of view can be placed outside of the car. A modification of 0.1 will yield a noticeable change. Head Offset: affects the position of the head-cam (cockpit view).Adjust FOV for: selects the in-game camera or cameras you want to edit.Vertical FOV: a larger number will translate into a wider field of view.Repeat until you're happy with the result. In DiRT 2, start a new race to test your settings.Tweak the values you previously entered and click Start. Click Restore to restore the default settings.Exit the race and Alt+ Tab ↹ to Dirt3FovChange. ![]() If you want to further adjust the settings:
Man, I am getting tired of doing that.Īnyway, during Early Access, Free Lives will continue to update the game with additional features, game modes, challenges, and gear which can be customized for your digital Johnson.įor more information on Devolver's latest bout of crazy, hit up the links. Genital Jousting isn’t exactly the newest game on the block. ![]() See what I mean? My inner juvenile is ranging to come out at this point. See full PC system requirements, expert reviews, user ratings, how to download Genital Jousting. Release Date: Summary Critic Reviews User Reviews Details & Credits Trailers & Videos Metascore 68 Mixed or average reviews based on 15 Critic Reviews Whats this User Score 8. if genital jousting got switch release Because Big is totally in Smash Bros. Players control a detached penis complete with testicles and an anus in a variety of game modes, each featuring a stimulating orgy of objectives: Penetrate and be penetrated as fast as possible or compete in absurd, silly and sexually suggestive games and challenges." Not really expecting much Switch to be mentioned (if at all), but theyre a. There are several moments that seem like legitimate choices to be made, only for those choices to be illusory a narrative device. No need to beat around the bush, this one is rock solid. Released originally in early access on November 17, 2016, the game was a party game based around flaccid penises (penii) penetrating. Genital Jousting is a pleasurable and girthy package with loads of fun mini-games to play with your friends that includes a short, funny yet impactful story mode that dares to touch on subjects other games won’t that will surely arouse some controversy. a disturbing mashup of Genital Jousting and that human centipede horror. The Legend of Zelda Tears of the Kingdom Nintendo Switch. Genital Jousting really doesn’t care for the player’s input. Genital Jousting isn’t exactly the newest game on the block. The game will be launching on Switch, as well as PC and Mac via the Epic Game. "Genital Jousting is an online and local multiplayer party game about flaccid penises and wiggly anuses for up to eight players at once. Its a digital game code that allows you to download Genital Jousting directly to your PC from. I wouldn't be able do it justice by summarizing it for you. It is up on Steam Early Access, and will run you $4.99.īelow is the blurb straight from Devolver. Titled Genital Jousting, Free Lives and Developer Digital announced the mind-boggling and shake your head funny game today. Traditional and Party Modes: Dabble in the throbbingly fast-paced Traditional Mode or jostle your way through some stiff competition in Party Mode including challenges like Double Delight, Obstacle Intercourse, and Weiner Round-Up.The latest multiplayer game from the folks who brought you Broforce is full of dicks. Try to switch to a wired connection instead of a wireless one. Online and Local Multiplayer for Eight: Genital Jousting is more fun with friends and can satisfy up to eight players at once! Find a few willing partners, cuddle up on the couch, and share controllers, or go online to joust with anonymous players from all over the world. You can do this by using an ethernet cable and connect it to your router. or may not be appropriate for viewing at work. If this is something that makes you uncomfortable or sounds unappealing, please do not purchase or play Genital Jousting, or even continue reading. What controllers or keyboards can play this game :: Genital Jousting General Discussions. And with your help, that's exactly what he'll do. He wants to show everyone his astounding rigidity. Due to the game's graphic depictions of 'sausages, zucchinis, eggplants, bananas' and the actual penetrations in someone else's 'doughnuts', this game. If you’ve no chance to play this with a group of your possibly intoxicated peers. Genital Jousting is very much an early access title. ![]() Play as John, a dick who wants to find a date for his high school reunion. Nintendo would not allowed to let Devolver Digital and/or Free Lives to port the game on Nintendo Switch because of their strict Seal of Approval policy. Every new mode is chaotic, fast-paced and full of genitals. ![]() Multiple game modes provide a stimulating orgy of objectives: penetrate and be penetrated as fast as possible or compete in absurd, silly, and sexually suggestive games and challenges. Players control a detached penis complete with testicles and an anus. Genital Jousting is an online and local multiplayer party game about flaccid penises and wiggly anuses for up to eight players at once. ![]() However, I might make some alternate versions of the mod that include different pricing models for those who would prefer more expensive potions. I can see why some people would feel that the prices are too low, but personally I'm happy with them as they are. Marioface5: Thanks for your feedback! Unfortunately, with the way the mod works (simply using effects that already existed in GUTS) I don't think I can check to prevent a potion from working if it would put the stat below zero.ĮDIT: I had multiple paragraphs about the price written here, it was kind of a stream of consciousness thing, but let me replace that with something more concise. I hope you enjoy the mod! If you experience any problems with it, please let me know and I'll try to look into it. ![]() That said, as always, you should BACK UP YOUR SAVE FOLDER before installing this mod as a precautionary measure. I've done my best to test this mod properly, and everything has worked as intended. There have been mods that do basically the same thing before, but since none of them were available on Nexus Mods I decided to create my own so that everyone here would have access to it. They should all be available at any General Goods merchant. The respec potion refunds all of your skill points, and the restat potions refund X amount of points from Y stat. ![]() One full respec potion, one single point restat potion per stat, one five point restat potion per stat, and one ten point restat potion per stat. Hello, everyone! I've just finished a new mod to share with you all: Respec and Restat Potions. Geometry Wars: Retro Evolved, developed by Bizarre Creations is the first main stand-alone game in the series. The soundtrack featured in G eometry Wars was composed by Chris Chudley from Audioantics, up until Geometry Wars 3: Dimensions, where he was replaced by Chris Mann of Mannmusic. The series is well known for its unforgiving nature and high difficulty, enemies reach higher numbers the longer the game lasts, and the ship that the player commands needs to be maneuvered with utmost precision since even touching an enemy means death. The game takes place on a rectangular playfield and the player controls a claw-shaped "ship" that can move in any direction using the left thumbstick, and can fire in any direction independently using the right thumbstick. The objective of Geometry Wars is to survive as long as possible and score as many points as possible by destroying an ever-increasing swarm of enemies. Geometry Wars is a series of Multidirectional Shooters, first appearing as a small minigame in the racing game Project Gotham Racing 2, and later becoming its own full-fledged franchise. This is the game’s version of grinding, and it’s one of the best ways to earn three stars in a level much easier.The current logo, as it appears in Geometry Wars 3: Dimensions If you’re looking to get yourself some three-star levels and soar up the leaderboard, then you can aim for a Quick Retry and play a stage you’ve already completed. Yes, black holes can be of help to you, but try not to shoot at it too much and make it go too fast for your own good. An active black hole would then suck in several enemies, and will keep sucking in more as it spins faster. You will encounter black holes occasionally in Geometry Wars 3: Dimensions, so how do you deal with them? Actually, these holes have a bluish hue to them (they aren’t entirely black holes, mind you), and they come about when you shoot a handful of bullets in to them. Don’t Shoot Too Much At Those Black Holes Connected red shapes can pop up in the middle of a tough battle, but don’t panic – just fly through them and keep firing at the enemy. As shooting these shapes won’t harm them, what you have to do is fly through them – you will get some green geoms for your trouble. Watch Out For The Connected Red ShapesĬonnected red shapes can be particularly tough customers – these are red shapes that have a white line connecting them. Movement is key to survival, and staying in one place will only end your game earlier than it should. Shoot in the direction your ship is traveling in, and always dodge enemies proactively. But since this is a strategy guide for newbies, we’ll stress this as one of the more important tips to share to you – you’ve got to keep on moving if you want to stay alive. If you’ve played previous Geometry Wars titles in the past, then you probably know this. ![]() This will allow you to increase your score substantially by jacking up your multiplier. Collect Green Geoms As You Shoot Down Your EnemiesĪs a rule of thumb, the main thing in Geometry Wars 3: Dimensions, or one of the main things, is to kill your enemies and collect the green geoms (no, that’s not a typo) that they leave in their wake. This arcade shooter game can be quite complex if you’re just starting out, but we’ve collected some useful Geometry Wars 3: Dimensions cheats, tips and strategies to help you start off the right way. ![]() There are also a good ten battle modes to choose from, five variants of companion drones, five special abilities, and a whole lot more, regardless whether you choose to play solo, against your friends, or against players from all over the world. There are 50 “Adventure” levels, plus five Classic Arcade levels, and two exclusive bonus levels for iOS gamers. Geometry Wars 3: Dimensions is the latest chapter in Activision’s popular series that has you encountering various 3D grid shapes, playing in different game modes, and fighting all manner of tough bosses. ![]() Incremental, concurrent, and real-time garbage collectors address these problems, with varying trade-offs. Unpredictable stalls can be unacceptable in real-time environments, in transaction processing, or in interactive programs. ![]() The moment when the garbage is actually collected can be unpredictable, resulting in stalls (pauses to shift/free memory) scattered throughout a session. The impact on performance was given by Apple as a reason for not adopting garbage collection in iOS, despite it being the most desired feature. Interaction with memory hierarchy effects can make this overhead intolerable in circumstances that are hard to predict or to detect in routine testing. The comparison however is made to a program generated by inserting deallocation calls using an oracle, implemented by collecting traces from programs run under a profiler, and the program is only correct for one particular execution of the program. A peer-reviewed paper from 2005 concluded that GC needs five times the memory to compensate for this overhead and to perform as fast as the same program using idealised explicit memory management. Therefore, the penalty for the convenience of not annotating object lifetime manually in the source code is overhead, which can impair program performance. GC uses computing resources to decide which memory to free.
She will meet Ben Tennyson in Ben 10's Adventures of My Little Pony: Friendship is Magic.She is close friends with Prairie Dusty Trails, Toffee Cocoa Cuddles, Berry Jars 'N' Jam and Sunny Side Up.Contents 1 Applejack's immediate family 1. The Apple family are the original founders of Ponyville. They are a large family of Earth pony farmers, who own apple farms all over Equestria. 4: Evolves from F AJACK - lvl 16 - Filly Applejack Evolves to G AJACK. The Apple family, sometimes called the Apple clan, note 1 are Applejack 's relatives in My Little Pony Friendship is Magic. While she guest stars, she wants to keep Apple Bloom safe from danger and villains because she does not let her become independent until she lets Apple Bloom to become independent. Applejack ChewThere are different reasons for a person to chew on their tongue. She will meet Ryan F-Freeman, Meg Griffin and their team in Ryan F-Freeman's Adventures of My Little Pony: Friendship is Magic.She will be best friends with Prairie Dusty Trails, Berry Jars 'N' Jam and Sunny Side Up in Jewel Sparkles' Adventure.She will appear in Jewel Sparkles' Adventure.Applejack met Connor Lacey and his friends in The Irelanders' Adventures of My Little Pony: Friendship is Magic (Full Movie).Applejack will join Barney and friends in Emerl's Adventures Of Barney in Outer Space.In Power Rangers Super Megaforce (TeamAdventureRules version), in her human form, she will become the Megaforce/Super Megaforce Yellow Ranger, taking over for Carmelita Fox.Applejack is also a good friend to Sandy Cheeks, Locomotive 131, Rabbit, Tummi Gummi, Cooki, Joe Kido and Gomamon.Applejack is a great friend to Kick Buttowski (who looks up to her as a sister), Gunther Magnuson, Darwin Watterson, and Gumball Watterson.She also owns a double barrel coach Shotgun.She as well as the rest of the Mane 6 are friends with Nanako Dojima in Yu Narukami and Aigis's adventures of My Little Pony: Friendship is Magic TV series, which is before Yu Narukami and Aigis's adventures of My Little Pony: Friendship is Magic-A Canterlot Wedding.In Stuington and Hiatt Grey's series, Applejack is Toby's girlfriend.Applejack will make occassional guest appearances in The FT Squad's Adventures With Phineas and Ferb, and will officially guest star in The FT Squad's Adventures With Phineas and Ferb: Summer Belongs to You.Applejack guest starred in The FT Squad Meets the Hunchback of Notre Dame and later will in The FT Squad's Adventures with the Winx Club.In this series, Louis the alligator shares a spot with her as the element of honesty. Applejack starred in The FT Squad's Adventures in My Little Pony: Friendship is Magic.(SPECIFICALLY CHAPTER 9) IF YOU HAVEN’T READ THAT YET, GO AWAY FROM THIS PAGE. Applejack will guest star in Pooh's Adventures of Soul Surfer. Excerpt of description by StarryOak from source: MASSIVE SPOILERS FOR THE IRONY OF APPLEJACK.Applejack met Ash, Littlefoot, Simba, Sora, Mickey, Spongebob, Alex, Tai and their friends in Pooh's Adventures of Thomas & Friends: Misty Island Rescue.Applejack met Pooh and his friends in Pooh's Adventures of My Little Pony: Friendship is Magic (Full Movie).She's also one of Cassidy the Pelican's closest friends, since they both wear western style clothing and got along perfectly when they first met. Applejack as a Filly Light-orange fur with white freckles on her cheeks. She represents the element of honesty in the elements of harmony. Despite this, she's kind and considerate. She's been known to be headstrong and insist on doing things by herself since she's very hard working. She's one of Twilight Sparkle's best friends. She's a very hard working pony who works as a farmer on Sweet Apple Acres in Ponyville with her grandmother Granny Smith, big brother Big McIntosh, little sister Apple Bloom and dog Winona. Do not hesitate to communicate with me.Applejack is an orange earth pony and one of the main characters in The FT Squad's Adventures in My Little Pony: Friendship is Magic. If there is a problem with your order or if you have questions or special requests about an item you are looking for. If you need additional details, feel free to send me a private message. It is important to carefully observe the photos which will describe the condition of the object. However, there may be signs of wear such as small stains, abrasions, strewn light hair, missing or altered parts. Thank you for visiting my shop because it is a privilege to receive you there.Įvery item in my shop has been reviewed. Let us drift into this colorful and imaginative dream world.Ī splendid gift for collectors, nostalgics and toy lovers. ![]() These dolls in the shape of a unicorn and a pony with their hair and tail that can be styled with their matching little comb and ribbon. Magnificent Fillies and Unicorns my little Pony G1 G2 G3 G4 Vintage Generation from several eras and years. * Sync data - Use Google Fit to sync meditation minutes from focus timer and study timer like plantie, flora green focus, focus to do, timetree, calm, study tracker, headspace, forest app, habitica, nox ocean, plant nanny, eggzy, flipd, flip, socratic, planty, study bunny, tomato timer, pomodoro timer, be focused, studysmarter, focus keeper, fatty cat, zoom, study blue, study buddy, saplingy, bft bear focus timer, sleep town, yeolpumta, my study life, study island, platie, donut dog, forrest, notion, pomofocus, finch, emphasis, flat tomato, virtual cottage, pomodoro technique, aloe bud, lifeat, focus mate, treedom, offtime, study planner, time tree, focos, study with me, tomatodo, ecosia, fishcure, trello, productive, mindly, planta, endel, ypt, and etc. * Pomodoro timer (tomato timer) - Give work productivity a boost, improve concentration and treat ADHD It also supports syncing Google Fit's meditation minutes from study timer like plantie, flora green focus, focus to do, timetree, calm, study tracker, headspace, forest app, habitica, nox ocean, plant nanny, eggzy, flipd, flip, socratic, planty, study bunny, tomato timer, pomodoro timer, be focused, studysmarter, focus keeper, fatty cat, zoom, study blue, study buddy, saplingy, bft bear focus timer, sleep town, yeolpumta, my study life, study island, platie, donut dog, forrest, notion, pomofocus, finch, emphasis, flat tomato, virtual cottage, pomodoro technique, aloe bud, lifeat, focus mate, treedom, offtime, study planner, time tree, focos, study with me, tomatodo, ecosia, fishcure, trello, productive, mindly, planta, endel, ypt, and etc. * Sync data from Focus Plant - In Focus Plant, you can use a countdown timer or a Pomodoro timer (tomato timer). ![]() * Meditation - You can also use the app as meditation gamification along with ADHD treatment * Rewards - Gamified tasks and rewards help you stay on track of your goals and to do * Records - Stay on track on your to do tasks to help with work productivity and self-control * Save the world - Fight off monsters and bring time order back to Focusland * Hundred of stages - Battle your way through various stages * Forge gears - Collect gear materials to forge powerful gears * Train hero - Use resources to train your hero * Stay focused - Collect hero resources when you put down the phone and manage life with ADHD ![]() * A unique game to beat phone addiction, help with ADHD, improve productivity and work concentration Overall, Focus Quest's unique blend of gamification and productivity tools offers users an engaging and effective way to boost their productivity and stay "centered" on their goals. By completing challenges and earning rewards, users can feel more "centered" and in control of their daily routines and long-term goals. It helps you to beat phone addiction, manage ADHD, and improve work productivity, self-control, and concentration.įocus Quest incorporates gamification elements to provide a "focus booster" for users seeking to overcome distractions and stay on task. ![]() Stay focused by pomodoro focus timer, study focus games, productivity timer appsįocus Quest is a productivity RPG gamification. ![]() So, the idea with laser cooling in this particular experiment is if I have atoms that are moving quickly towards my laser, they absorb photons and get slowed down, so I can slow down the fast ones and not mess with the ones that are already slow enough. Absorption of the light is dependent on how fast the atom is moving. Now, light carries momentum, and when an atom absorbs one of its preferred photons, it gets a kick from the momentum of the photon, sort of like catching a heavy medicine ball if you’re standing on ice. When I spoke to Jeffrey, I asked: Can you explain how the technique that you use, which is called laser cooling, works?Īntihydrogen, like other atoms as far as we know, can absorb light, and atoms like to absorb particular colours of light. Slowing down antihydrogen could unlock experiments that will be able to compare anti-hydrogen and regular hydrogen with unprecedented precision. Antihydrogen consists of just one antiproton and one antielectron. In an experiment described in this week’s Nature, Jeffrey and the team were investigating how to slow down the movements of the simplest possible anti-atom – antihydrogen. It’s certainly lucky for us that it did, since if there’d been equal amounts of both it all would have annihilated and there would be no stars, planets, humans or podcasts. If physicists found some deviation in antimatter’s properties from the mirror image of matter that we’d expect, that could explain this fundamental question – why our Universe was created with a surplus of matter over antimatter. ![]() Why did the Universe evolve to matter instead of antimatter? If the Universe started as energy and it went on to evolve some mass, what happened to the anti-mass that should have been created at the same time? And the answer is we simply don’t know. The answer is how could you not if you can? So, there’s some mysteries associated with these antimatter particles, and this dates back to the beginning of the Universe. So, given that these experiments are so challenging, why are Jeffrey and collaborators painstakingly studying antimatter? It means that any experiments with antimatter are incredibly tricky, as one wrong move and your antimatter bumps into some matter and is annihilated. The interesting thing about these particles is they don’t exist normally because if they come into contact with normal matter they will annihilate and release a bunch of energy. Now, we know in modern physics that for every particle that we’ve been able to observe, you can have an equal but opposite antiparticle. This is physicist Jeffrey Hangst of Aarhus University in Denmark, and spokesperson of the ALPHA antimatter experiment at CERN in Switzerland. You see, in physics, there’s also something called antimatter.įor the lay person, we like to think of antimatter as kind of an evil twin of the matter that makes up everything that you and I know. Our Universe is made up of matter, but matter is only half the story. I’m Nick Petrić Howe.įirst up on the show, reporter Adam Levy is here to talk about an important matter. This week, antimatter gets cool…Īnd the global cost of biological invasions. Head here for the Nature Podcast RSS feed. Never miss an episode: Subscribe to the Nature Podcast on Apple Podcasts, Google Podcasts, Spotify or your favourite podcast app. Subscribe to Nature Briefing, an unmissable daily round-up of science news, opinion and analysis free in your inbox every weekday. Science: Octopuses, like humans, sleep in two stages The Financial Times: The bank effect and the big boat blocking the Suez This time, the physics that might explain how a ship blocked the Suez Canal, and a new insight into octopuses’ sleep patterns. We discuss some highlights from the Nature Briefing. This week, researchers estimate that the economic impact of invasive species to be over US $1 trillion. These species can cause loss of biodiversity and a host of damage to their new environments. Invasive alien species are organisms that end up in places where they don’t really belong, usually as a result of human activity. Research Highlight: Rabbits that do ‘handstands’ help to find a gene for hopping 11:53 Cost of invasion Research Highlight: Rising temperatures spark boom in Arctic lightning News and Views: Antimatter cooled by laser light 09:27 Research HighlightsĪ dramatic increase in Arctic lightning strikes, and an acrobatic bunny helps researchers understand hopping. Now though, a team at CERN have developed a way to trap and cool antihydrogen atoms using lasers, allowing them to better study its properties. In this episode: 00:44 Cooling antimatter with a laser focusĪntimatter is annihilated whenever it interacts with regular matter, which makes it tough for physicists to investigate. ![]() Figures 1, 3, 5, 6, S2, and S3 were generated in Biorender. We would also like to thank Charina Fabilane for assistance in workflow optimization. The authors would like to thank Paul Sargunas for assistance with protein purifications. T1 - Discovery of antibodies targeting multipass transmembrane proteins using a suspension cell-based evolutionary approach We implemented our strategy to discover high-affinity antibodies against four distinct GPCR target proteins, demonstrating the potential for our cell-based screening workflow to advance the discovery of antibody therapeutics targeting membrane proteins.", Here, we developed suspension cell-based whole-cell panning methodologies to screen for specific binders against GPCRs within a naive yeast-displayed antibody library. These challenges, largely due to difficulties with recombinant expression of multipass transmembrane proteins, can be overcome using whole-cell screening techniques, which enable presentation of the functional antigen in its native conformation. However, major limitations exist in developing antibodies that recognize complex, multipass transmembrane proteins, such as G-protein-coupled receptors (GPCRs). We implemented our strategy to discover high-affinity antibodies against four distinct GPCR target proteins, demonstrating the potential for our cell-based screening workflow to advance the discovery of antibody therapeutics targeting membrane proteins.Ībstract = "Due to their critical functions in cell sensing and signal processing, membrane proteins are highly preferred as pharmacological targets, and antibody drugs constitute the fastest growing category of therapeutic agents on the pharmaceutical market. ![]() Due to their critical functions in cell sensing and signal processing, membrane proteins are highly preferred as pharmacological targets, and antibody drugs constitute the fastest growing category of therapeutic agents on the pharmaceutical market. |