Analysis of the V genes coding for a monospecific human antibody to myosin and functional expression of single chain Fv fragments.
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.
Laroche-Traineau
J
J
CNRS UMR 5533, Hôpital Cardiologique, Pessac, France. Jeanny.Laroche@umr5533.u-bordeaux2.fr
Jacobin
M J
MJ
Biard-Piechaczyk
M
M
Vuillemin
L
L
Chagnaud
J L
JL
Pau
B
B
Nurden
A T
AT
Clofent-Sanchez
G
G
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000235:genetics*; D001402:B-Lymphocytes / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli; D005786:Gene Expression Regulation; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007075:Immunoglobulin M / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008969:Molecular Sequence Data; D009206:Myocardium / Q000276:immunology; D018995:Myosin Heavy Chains / Q000276:immunology*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007075:Immunoglobulin M; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D018995:Myosin Heavy Chains