Binding of monomeric and aggregated immunoglobulin to enzymes. A source of artefact in antibody assays.
Pepsinogen has previously been shown to bind non-specifically to immune complexes and aggregated immunoglobulins. We demonstrate here using a solid-phase immunoassay that immunoglobulins aggregated by heat or glutaraldehyde bind non-specifically to several different enzymes. Some of these, including pepsinogen (marketed as pepsin), hyaluronidase and trypsin, are used in the breakdown of tissues or biochemical preparations during the preparation of antigens. Contamination of impure antigens by enzyme is likely to lead to products which bind non-specifically to immune complexes. This can cause misidentification of complexes as antibodies. We recommend that all tests for specific antibody involving the use of antigens prepared by these or other enzymes should include a control with aggregated immunoglobulin substituted for the test serum.
O'Hara
B P
BP
ARC Bone and Joint Research Unit, London Hospital Medical College, U.K.
Pyle
J
J
McCarthy
D
D
Archer
J R
JR
D000906:Antibodies / Q000032:analysis*; D000936:Antigen-Antibody Complex; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004798:Enzymes; D006358:Hot Temperature; D006801:Humans; D007136:Immunoglobulins; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D010435:Pepsinogens; D011485:Protein Binding
D000906:Antibodies; D000936:Antigen-Antibody Complex; D004798:Enzymes; D007136:Immunoglobulins; D046911:Macromolecular Substances; D010435:Pepsinogens