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Comparison of two functional kappa light‐chain transcripts amplified from a hybridoma
Author(s) -
Yang Juan,
Zhu Huifen,
Tan Zheng,
He Fengrong,
Sun Xiaoxu,
Hong Yi,
Hu Heyu,
Bian Jing,
Lin Yu,
Lei Ping,
Shen Guanxin
Publication year - 2013
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1080
Subject(s) - immunoglobulin light chain , somatic hypermutation , biology , monoclonal antibody , microbiology and biotechnology , kappa , phylogenetic tree , antibody , genetics , functional analysis , computational biology , gene , b cell , linguistics , philosophy
Three heavy‐chain and three kappa (κ)‐chain transcripts were amplified from hybridoma cells secreting a monoclonal antibody (m A b) against transferrin receptor. Sequence analysis via IMGT / V ‐ QUEST yielded the functional/aberrant prediction. Two functional κ‐chain transcripts, V κ2 and V κ3, and one functional V H 1 were revealed. Comprehensive bioinformatics analyses including sequence alignment, phylogenetic tree, somatic hypermutation prediction, and three‐dimensional‐molecular structure modeling were used to predict the origin of the two κ‐chain transcripts. The results of bioinformatics analysis suggest that V κ3 is derived from the myeloma partner of the hybridoma; V κ2 is derived from B‐cell. Functional transcripts V H 1 and V κ2 and V κ3 were then used to construct two chimeric antibodies chi‐ C 2 ( V κ2– V H 1) and chi‐ C 3 ( V κ3–V H 1), respectively. Antigen‐binding experiments showed that only chi‐ C 2 remained the same affinity as its parental mAb. Possible explanations for the coexistence of two functional κ‐chain transcripts and the different affinity of the two chimeric antibodies are discussed.