Generating a panel of highly specific antibodies to 20 human SH2 domains by phage display.
Generating a panel of highly specific antibodies to 20 human SH2 domains by phage display.
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DOI：
10.1093/protein/gzq003
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发表时间：
2010-04
期刊：
影响因子：
--
通讯作者：
McCafferty J
中科院分区：
文献类型：
--
作者：
Pershad K;Pavlovic JD;Gräslund S;Nilsson P;Colwill K;Karatt-Vellatt A;Schofield DJ;Dyson MR;Pawson T;Kay BK;McCafferty J
To demonstrate the utility of phage display in generating highly specific antibodies, affinity selections were conducted on 20 related Src Homology 2 (SH2) domains (ABL1, ABL2, BTK, BCAR3, CRK, FYN, GRB2, GRAP2, LYN, LCK, NCK1, PTPN11 C, PIK3R1 C, PLCγ1 C, RASA1 C, SHC1, SH2D1A, SYK N, VAV1 and the tandem domains of ZAP70). The domains were expressed in Escherichia coli, purified and used in affinity selection experiments. In total, 1292/3800 of the resultant antibodies were shown to bind the target antigen. Of the 695 further evaluated in specificity ELISAs against all 20 SH2 domains, 379 antibodies were identified with unique specificity (i.e. monospecific). Sequence analysis revealed that there were at least 150 different clones with 1–19 different antibodies/antigen. This includes antibodies that distinguish between ABL1 and ABL2, despite their 89% sequence identity. Specificity was confirmed for many on protein arrays fabricated with 432 different proteins. Thus, even though the SH2 domains share a common three-dimensional structure and 20–89% identity at the primary structure level, we were able to isolate antibodies with exquisite specificity within this family of structurally related domains.
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DOI：
10.1073/pnas.95.11.6157
发表时间：
1998-05-26
影响因子：
11.1
作者：
Sheets, MD;Amersdorfer, P;Marks, JD
通讯作者：
Marks, JD
将噬菌体显示到高吞吐抗体的产生和表征。
影响因子：
12.3
作者：
Schofield, Darren J;Pope, Anthony R;Clementel, Veronica;Buckell, Jenny;Chapple, Susan Dj;Clarke, Kay F;Conquer, Jennie S;Crofts, Anna M;Crowther, Sandra R E;Dyson, Michael R;Flack, Gillian;Griffin, Gareth J;Hooks, Yvette;Howat, William J;Kolb-Kokocinski, Anja;Kunze, Susan;Martin, Cecile D;Maslen, Gareth L;Mitchell, Joanne N;O'Sullivan, Maureen;Perera, Rajika L;Roake, Wendy;Shadbolt, S Paul;Vincent, Karen J;Warford, Anthony;Wilson, Wendy E;Xie, Jane;Young, Joyce L;McCafferty, John
通讯作者：
McCafferty, John
影响因子：
3.4
作者：
Nilsson, P;Paavilainen, L;Uhlen, M
通讯作者：
Uhlen, M
影响因子：
7
作者：
Huang, Haiming;Li, Lei;Li, Shawn S. -C.
通讯作者：
Li, Shawn S. -C.
影响因子：
48
作者：
Taussig, Michael J.;Stoevesandt, Oda;Uhlen, Mathias
通讯作者：
Uhlen, Mathias