Barnase-barstar high affinity interaction phenomenon as the base for the heterogenous bioluminescence pseudorabies virus' immunoassay.
The effective new variant of "sandwich" bioluminescent enzyme immunoassay (BEIA) for the sensitive detection of glycoprotein B (gB) of pseudorabies virus (PrV) was presently developed. The high affinity interaction of barnase-barstar protein pair and photoprotein obelin as bioluminescent marker were for the first time successfully applied to BEIA development. Preliminary the two monoclonal antibodies, 11/5 and 34/2, were raised against gB for ELISA PrV detection. Presently we used the same immuno-"sandwich" principle for BEIA. To do this the two different bioconjugates were elaborated. Recombinant barnase was chemically conjugated with monoclonal anti-PrV's gB IgG, and also barstar was fused in frame to obelin. The characteristics of BEIA method have been compared to ELISA PrV detection. We have shown the proposed here gB-BEIA was 40-fold more sensitive as opposed to gB-ELISA test. The construction might have a broad promise in multiple potential immunological applications.
Rudenko
Natalia V
NV
Branch of Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, RAS, 142290, Pushchino, Russia.
Sinegina
Lilia L
LL
Arzhanov
Maxim A
MA
Ksenzenko
Vladimir N
VN
Ivashina
Tatiana V
TV
Morenkov
Oleg S
OS
Shaloiko
Lyubov A
LA
Vinokurov
Leonid M
LM
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002460:Cell Line; D006224:Cricetinae; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D007118:Immunoassay / Q000379:methods; D049449:Luminescence; D008164:Luminescent Proteins / Q000235:genetics; D016366:Open Reading Frames; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; C010497:obelin; C057438:barstar protein, Bacillus amyloliquefaciens