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Identification of human antibody fragment clones specific for tetanus toxoid in a bacteriophage lambda immunoexpression library.

Abstract：

:We have applied a molecular biology approach to the identification of human monoclonal antibodies. Human peripheral blood lymphocyte mRNA was converted to cDNA and a select subset was amplified by the polymerase chain reaction. These products, containing coding sequences for numerous immunoglobulin heavy- and kappa light-chain variable and constant region domains, were inserted into modified bacteriophage lambda expression vectors and introduced into Escherichia coli by infection to yield a combinatorial immunoexpression library. Clones with binding activity to tetanus toxoid were identified by filter hybridization with radiolabeled antigen and appeared at a frequency of 0.2% in the library. These human antigen binding fragments, consisting of a heavy-chain fragment covalently linked to a light chain, displayed high affinity of binding to tetanus toxoid with equilibrium constants in the nanomolar range but did not cross-react with other proteins tested. We estimate that this human immunoexpression library contains 20,000 clones with high affinity and specificity to our chosen antigen.

journal_name

Proc Natl Acad Sci U S A

journal_title

Proceedings of the National Academy of Sciences of the United States of America

authors

Mullinax RL,Gross EA,Amberg JR,Hay BN,Hogrefe HH,Kubitz MM,Greener A,Alting-Mees M,Ardourel D,Short JM

doi

10.1073/pnas.87.20.8095

subject

Has Abstract,Author List Incomplete

pub_date

1990-10-01 00:00:00

pages

8095-9

issue

20

eissn

0027-8424

issn

1091-6490

journal_volume

87

pub_type

杂志文章

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