A species and organ specific glomerular basement membrane antigen

Abstract

By the use of a monoclonal antibody reacting with the human glomerular basement membrane exclusively we have been isolating a specific glomerular antigen. The antibody failed to react with other renal structures and other basement membranes and extracellular matrix constituents of other organs or plasma proteins. It did not react with glomerular basement membranes of other species as mice and rats. For the isolation of the antigen we applied affinity chromatography, on Sepharose beads coupled to the monoclonal antibody PM II 34 G3. From this column the antigen was eluted under acid conditions. In 8% polyacrylamide gel electrophoresis the approximate molecular weight was estimated with 14400 daltons, which was confirmed on high pressure liquid chromatography using the Bio-Sil TSK method. The antigen was temperature sensitive in that at high temperatures (60 degrees C) several bands on SDS-PAGE and several peaks on HPLC could be noted. This could be responsible for the crystal formation after treatment/concentration on the rotary vacuum pump at 60 degrees C. Preliminary data of amino acid analysis showed a high glycine content pointing towards a collageneous molecule. But cross reactivity with many connective tissue proteins could be ruled out by immuno-histochemical techniques and affinity chromatography. A major point of interest is that this antigen can be detected in human urine under physiological conditions. We are herewith reporting the first species and organ specific glomerular basement membrane antigen.