INTENDED USE

This HAMA ELISA is to be used for the quantitative determination of human anti-mouse antibody (HAMA) in serum. It is useful for the prognosis, monitoring and follow-up patients treated with mouse IgG. This kit is intended for LABORTAROY RESEARCH USE ONLY and is not for use in diagnostic or therapeutic procedures.

INTRODUCTION

Human Anti-Mouse Antibody (HAMA) are antibodies found in human serum which have the ability to bind to mouse immunoglobulin G (IgG). The presence of HAMA is the result of an immune reaction following an exposure to mice or other similar agent, which was able to induce the immune system to generate an antibody able to bind to mouse IgG. HAMA is commonly found in patients following treated with mouse monoclonal antibodies associated with some therapeutic or diagnostic procedures. Some human auto-antibodies, the most common of which is rheumatoid factor (Rf), by virtue of their cross reactivity are able to bind mouse IgG. The HAMA assay measures HAMA across a wide dynamic range and it is not necessary to predilute human serum samples before the HAMA ELISA testing.

PRINCIPLE OF THE ASSAY

This assay applies an enzyme-linked immunosorbent assay (ELISA) technique. A purified mouse IgG has been pre-coated onto the wells of microplate. Standard (rabbit anti-mouse IgG) and specimens are added into the appropriate wells simultaneously with a HRP-labelled mouse IgG, and incubated. The Human Anti-Mouse Antibody (HAMA), if present, will bind with the mouse IgG which coated on the wells and HRP-labelled mouse IgG as well, forming a "Sandwich". Following a wash to remove any unbound substance, a substrate solution is added. The developed colour is directly proportional to the anti-mouse antibody concentration in the specimen and Standard samples.

This ELISA Kit includes a Calibration Diluent for preparing a 2-fold dilution of Standard series. It allows the operator to produce a standard curve with Optical Density (O.D) versus Human Anti-Mouse Antibody concentration (ng/mL). The concentration of Human Anti-Mouse Antibody in the sample is then determined by comparing the O.D. of the sample to the standard curve.