Immunoglobulins of the snake Psammophis sibilans. Studies using a monoclonal antibody

A mouse monoclonal antibody (mAb) raised against serum immunoglobulins (Ig) of the snake. Psammophis sibilans stained in indirect immunofluorescence a proportion of snake splenic and peripheral blood lymphocytes, whereas it did not react with thymocytes, erythrocytes, brain, heart, lung, liver or kidney cells. The mAb, designated SR-2, combined in enzyme-linked immunosorbent assay (ELISA) with serum proteins of each of 20 individual P. sibilans tested. On Western blots of P. sibilans reduced whole serum proteins, purified Ig, or anti-rat erythrocyte (RRBC) antibodies eluted from glutaraldehyde-fixed RRBC, mAb SR-2 identified two bands of apparent molecular weight (m.w.) of 60,000 and 51,000 daltons. These bands were due to distinct polypeptides and not resulting from heterogeneous glycosylation of a single polypeptide, as they both were readily detected after periodate oxidation or endoglycosidase-F treatment of serum proteins and isolated Ig. MAb SR-2 bound to CNBr-activated Sepharose 4B precipitated from P. sibilans 125I-labeled serum proteins under non-reducing conditions a band that did not enter 7.5 or 9-16% gel and one of about 150,000 daltons. Under reducing conditions, two heavy bands of approximately 63,000 and 50,000 daltons and two light chains of apparent mass 23,000 and 20,000 bands were precipitated. The data presented provide, for the first time, substantial information on the molecular characteristics of snake Ig.