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Synthesis of Rh Fv phage‐antibodies using VH and VL germline genes
Author(s) -
HughesJones Nevin,
Bye Jacqueline,
Gorick Barbara,
Marks James,
Ouwehand Willem
Publication year - 1999
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1999.01412.x
Subject(s) - antibody , germline , immunoglobulin light chain , phage display , microbiology and biotechnology , antigen , gene , biology , chemistry , genetics
Antibodies to the D antigen of the Rh system use a restricted set of immunoglobulin V and J gene segments, especially V H DP50 and DP63, J H 6, V λ DPL16 and J λ 2/3. These gene segments may confer a natural affinity on the antibodies for the D antigen and this hypothesis has been tested by constructing two single‐chain Fv phage‐antibody libraries based on the germline gene segments DP50 and DP63; structural variability was obtained by insertion of 11 amino acids in random sequence in the V H CDR3. 10 anti‐D antibodies were selected from these libraries, each with a unique V H CDR3. In contrast, selections with the CcEe antigens produced antibodies reacting with the Rh polypeptide molecules but without strict blood group specificity. One of these latter DP50‐based antibodies was converted into 12 different antibodies with specificity for E by replacing the original germline light chain with chains from a rearranged L chain library. The CDR1 and CDR2 sequences of the DP50‐based antibodies were common to both anti‐D and anti‐E molecules; differentiation between D and E specificity was dependent on V H CDR3 sequences and their correct pairing with an appropriate L chain.