The use of beta-galactosidase fusion proteins encoding the early region 1 transforming proteins of adenovirus type 12 to examine the humoral response in tumour-bearing animals.
Sera from 26 rats bearing tumours induced by wild-type (wt) and mutant human adenovirus type 12 (Ad12), or by cells transformed with these viruses, were analysed for antibodies against the early region 1 (E1) transforming proteins. Six Ad12-beta-galactosidase fusion proteins encoding different regions of the Ad12 E1 proteins were constructed. The sera from the tumour-bearing animals reacted most strongly with the fusion protein encoding the N terminus of the E1A protein. Tumour-bearing rats exposed to the E1B 54K and 19K proteins showed strong reactions with the N terminus of the 54K protein and the C terminus of the 19K protein. Monospecific polyclonal antisera were raised against five of the fusion proteins by immunization of rats and rabbits; these sera cross-reacted with the purified native protein. No antibodies could be obtained which recognized a fusion protein containing amino acids 136 to 268 of the 54K protein. The fusion proteins were also used to purify monospecific antisera from tumour-bearer sera using affinity chromatography.
Merrick
R M
RM
Department of Cancer Studies, Medical School, University of Birmingham, U.K.
Grand
R J
RJ
Brown
J C
JC
Gallimore
P H
PH
D000257:Adenoviridae Infections / Q000276:immunology*; D017456:Adenovirus Early Proteins; D000260:Adenoviruses, Human / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000917:Antibody Formation; D015153:Blotting, Western; D002460:Cell Line; D007106:Immune Sera; D008970:Molecular Weight; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000276:immunology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000276:immunology; D015183:Restriction Mapping; D001616:beta-Galactosidase / Q000235:genetics*
D017456:Adenovirus Early Proteins; D007106:Immune Sera; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase