Premium
Analysis of the V genes coding for a monospecific human antibody to myosin and functional expression of single chain Fv fragments
Author(s) -
Laroche-Traineau Jeanny,
Jacobin Marie-Josée,
Biard-Piechaczyk Martine,
Vuillemin Luce,
Chagnaud Jean-Luc,
Pau Bernard,
Nurden Alan T.,
Clofent-Sanchez Gisèle
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01308-3
Subject(s) - monoclonal antibody , microbiology and biotechnology , myosin , antigen , immunoglobulin light chain , antibody , escherichia coli , biology , gene , chemistry , genetics
A monospecific human IgM monoclonal antibody (mAb), reactive with myosin from human heart, has been obtained by EBV transformation. This mAb may have a diagnostic potential in the imaging of myocardial necrosis. However, owing to the fact that the molecular mass of an IgM is 900 kDa, a poor diffusion and a slow penetration inside necrotic myocytes could reduce its capacity for scintigraphic detection. In order to alleviate these problems, we constructed the scFv by cloning the VH and VL domains into the pHOG21 vector. Analysis of the V genes proved an unmutated configuration showing that the immortalized B cell issued from the primary IgM repertoire. The expression product in Escherichia coli was a 35 kDa scFv fragment with the antigen‐binding specificity of the parental mAb.