Immunoglobulin V regions of a bactericidal anti-Neisseria meningitidis outer membrane protein monoclonal antibody.
C6 is a potentially therapeutic murine monoclonal antibody that recognizes the class 1 outer membrane protein of Neisseria meningitidis. C6 specifically immunoblots this antigen and augments in vitro killing of N. meningitidis bacteria. We describe a general method of obtaining the heavy and light chain variable-region sequence from immunoglobulin-secreting cells. The method uses mixed polymerase chain reaction (PCR) primers designed from the 5' end of the framework 1 (FR1) sequences of the heavy and light chains, and 3'-end primers for constant-region conserved sequences. The method has been applied to the cloning and sequencing of the variable region of C6 to construct a humanized monoclonal antibody. Rapid amplification and sequencing of variable regions by this general method have multiple applications in the study of the immune response to infectious diseases.
Larrick
J W
JW
Genelabs Inc., Redwood City, California.
Coloma
M J
MJ
del Valle
J
J
Fernandez
M E
ME
Fry
K E
KE
Gavilondo-Cowley
J V
JV
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000235:genetics* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology*; D001483:Base Sequence; D004269:DNA, Bacterial; D005784:Gene Amplification; D015151:Immunoblotting; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000276:immunology*; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region