副镜
免疫球蛋白轻链
生物
寡核苷酸
基因
分子生物学
基因家族
遗传学
免疫球蛋白重链
抗体
DNA测序
底漆(化妆品)
单克隆抗体
基因组
化学
有机化学
作者
Thierry Chardès,Sylvie Villard,Gaëlle Ferrières,Martine Piechaczyk,Martine Cérutti,G. Devauchelle,Bernard Pau
出处
期刊:FEBS Letters
[Wiley]
日期:1999-06-11
卷期号:452 (3): 386-394
被引量:30
标识
DOI:10.1016/s0014-5793(99)00649-3
摘要
We have designed two original sets of oligonucleotide primers hybridizing the relatively conserved motifs within the immunoglobulin signal sequences of each of the 15 heavy chain and 18 kappa light chain gene families. Comparison of these 5′ primers with the immunoglobulin signal sequences referenced in the Kabat database suggests that these oligonucleotide primers should hybridize with 89.4% of the 428 mouse heavy chain signal sequences and with 91.8% of the 320 kappa light chain signal sequences with no mismatch. Following PCR amplification using the designed primers and direct sequencing of the amplified products, we obtained full‐length variable sequences belonging to major (V H 1, V H 2, V H 3, Vκ1 and Vκ21) but also small‐sized (V H 9, V H 14, Vκ2, Vκ9A/9B, Vκ12/13, Vκ23 and Vκ33/34) gene families, from nine murine monoclonal antibodies. This strategy could be a powerful tool for antibody sequence assessment whatever the V gene family before humanization of mouse monoclonal antibody or identification of paratope‐derived peptides.
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