Isolation and partial characterization of catalytic antibodies with oligonuclease activity from bovine colostrum.
Catalytic antibodies (abzymes) which hydrolyze RNA and DNA were isolated from bovine colostrum by sequential chromatography on Protein A Sepharose, denaturated DNA-cellulose, Mono Q, and gel permeation chromatography on Superose 12 at pH 2.3 after acidic shock. Metachromatic agar containing toluidine blue and yeast RNA was used to measure RNase activity. Electrophoresis in agarose showed DNase activity on plasmid DNA from Escherichia coli and DNA from calf thymus in fractions from all 4 purification steps. Gel permeation chromatography showed that the abzymes hydrolysed both a single-stranded polyadenylic acid (Poly A) and single-stranded polycitidylic acid (Poly C), while partially purified RNase from the colostrum hydrolysed Poly (C), but not Poly (A). Electrophoresis of purified abzymes under denaturing conditions showed protein bands of molecular mass corresponding to heavy and light chains of IgG. The abzymes immunoreacted with anti-bovine IgG. The RNase activity of the purified abzymes represented 0.022% of total RNase activity in the colostrum; acid shock and gel filtration at low pH reduced the specific RNase activity of abzymes 3.6-fold. The RNase activity of abzymes at pH 6.6 was reduced by 90% by heat treatment at 75 degrees C for 52 min.
Stepaniak
Leszek
L
Department of Food Science, Agricultural University of Norway, As.
D000818:Animals; D017151:Antibodies, Catalytic / Q000302:isolation & purification* / Q000378:metabolism*; D002417:Cattle; D002850:Chromatography, Gel; D003126:Colostrum / Q000201:enzymology* / Q000276:immunology*; D003851:Deoxyribonucleases / Q000302:isolation & purification* / Q000378:metabolism*; D004926:Escherichia coli; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008970:Molecular Weight; D009841:Oligonucleotides / Q000378:metabolism; D012260:Ribonucleases / Q000302:isolation & purification* / Q000378:metabolism*
D017151:Antibodies, Catalytic; D009841:Oligonucleotides; D003851:Deoxyribonucleases; D012260:Ribonucleases