Affinity screening for weak monoclonal antibodies

When selecting for monoclonal antibodies of a desired affinity, affinity chromatography can be a feasible alternative. This is of particular interest when low affinity monoclonal antibodies (dissociation constant (Kd) > 10(-4) M) are screened, as they are not easily recognised by traditional immunoassay procedures. In this study we have evaluated this approach by monitoring low affinity monoclonal antibodies on high performance liquid affinity chromatography columns with oligosaccharides, dinitrophenol and digoxin as immobilised antigen. Crude monoclonal antibodies in ascites or cell culture supematants, directed against these antigens, were retarded or adsorbed according to affinity or avidity on the antigen columns. Based on antibody retention, we were able to select hybridomas with the desired low affinity characteristics.

MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Immunologi inom det medicinska området hsv//swe
MEDICAL AND HEALTH SCIENCES Basic Medicine Immunology in the medical area hsv//eng
Animals
Antibodies, Monoclonal/immunology
Antibody Affinity
Ascitic Fluid/immunology
Chromatography, Affinity
Chromatography, High Pressure Liquid
Digoxin/immunology
Dinitrophenols/immunology
Hemocyanins/immunology
Immunoglobulin M/immunology
Mice
Oligosaccharides/immunology

Grubb, AndersLund University,Lunds universitet,Avdelningen för klinisk kemi och farmakologi,Institutionen för laboratoriemedicin,Medicinska fakulteten,Cystatin C, njursjukdom, amyloidos och antibiotika,Forskargrupper vid Lunds universitet,Division of Clinical Chemistry and Pharmacology,Department of Laboratory Medicine,Faculty of Medicine,Cystatin C, renal disease, amyloidosis and antibiotics,Lund University Research Groups,Skåne University Hospital(Swepub:lu)kkem-agr (författare)
Ohlson, StenUniversity of Kalmar (författare)
University of KalmarAvdelningen för klinisk kemi och farmakologi (creator_code:org_t)

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