PiggyBac Antibody Heav做水y Chain Gene Expression Vector

Recombinant antibod裡線ies have extensive application志中s in the fields of diagnosti要通cs and therapeu視錯tic medicine. Curren銀街tly, there are 舊睡about 180 monoclonal 飛到antibody products approve不冷d worldwide targeting specific 通呢proteins, such as P是雪D-L1, HER2, IL-17R, and VEGF. The d要視evelopment of gen又校etic engineering has facilitated suc生報cessful production of the機時se therapeutic antibodies, which hav西村e ideal tumor penetration behav飛員ior, short retention times, and re要金duced immunogenicity. These the雜有rapeutic recombinant an海空tibodies are mainly produced i車光n mammalian cells in vitro (是風e.g., CHO cells),鄉小 since the post-transcription用子al modification in this ex訊微pression system is clos讀議est to the in vivo processes輛黑 in humans. 

The antigen-bin年上ding site of all antibodi校們es consists of VL+章我VH dimers where VL and VH represen視了t variable fragment 錢線light chains and heavy chains, respe書亮ctively. Within these fragments, 子花most sequence var黃在iation is located in co爸上mplementarity-determini務報ng regions (CDRs), which influence笑低s antigen-binding specificities. By cus從討tomizing sequences 車快encoding the heavy chain vari長議able region, customers是見 may generate vectors to satisfy the從多ir personalized ne爸個ed for antigen-binding specificities. I放化n addition to the variable (V) region,吃光 the heavy and light c雜鄉hains contain a co費對nstant (C) region, wh北煙ich determines the antibody’s is外民otype. Of the five immunoglobulin (an計森tibody) isotypes (IgG, IgA, IgM, IgD a一廠nd IgE), IgG is the most abun關視dant in circulation. More朋章over, almost all backbones o小我f the approved ther放生apeutic antibodies 子吧are IgG. In the vector desig東店n process, the heavy cha你問in constant region may著什 either be selected from ou雪金r popular heavy chain c習他onstant region database 子計(containing human and mouse IgG1 市學and mouse IgG2a and 2b) o但就r be pasted by cus離可tomers using their own sequen站厭ce. Additionally, signal peptides 綠我(e.g., IL-2 sig) can be adde生兒d into the vector to increas輛間e secretion of the recombinant a體化ntibody.

VectorBuilder’s piggyBac antibody hea通去vy chain expression 北城vector system is a highly efficient風機 tool for achieving non-v件放iral, transposon-b廠西ased delivery of anti得舊body heavy chain expression cass報習ette into mammalian cells. This s科如ystem contains two vectors, both of動呢 which are engineered as E. coli pla農作smids. The first照女 vector is referred to as坐視 the helper PBase plasmid, encodi視靜ng the transposase. The second 微是vector is referre業冷d to as the piggyBac transposon 我弟plasmid which contains討個 two terminal re老黑peats (TRs) bracketing the antibo行件dy heavy chain 請音expression cassette to be 不書transposed. When the helper PBa視來se plasmid and the piggy會場Bac transposon plasmids are co-trans慢嗎fected into target cells, the transposa紙花se from the helper plasmid will reco劇內gnize the two TRs on the transposon又空 and insert the flanked antibo嗎訊dy heavy chain expre白能ssion cassette into the host geno技對me. Insertion typically occurs a畫從t host chromosomal sites th校她at contain the 去哥TTAA sequence, which is duplicated on t樹路he two flanks of the integrated fragme器關nt. PiggyBac is a cla站綠ss II transposo的媽n, meaning that it 校睡moves in a cut-and-paste ma動區nner, hopping from 拍師place to place wit近自hout leaving copi些兒es behind. (In contrast, class腦都 I transposons move in co弟線py-and-paste manner). Because th短關e helper plasmid機有 is only transiently tra吧答nsfected into host cells, it will get 黃風lost over time. With t木民he loss of the helper plasmid愛是, the integration of genes encoding t放筆he antibody heavy chain b都機ecomes permanent in the geno風我me of host cells.

For further information about 老黃this vector system, please ref坐數er to the papers below.