PRepaRation and Identification of Monoclonal Antibody of PRotein &sigma;3 fRom Avian ReoviRus

doi:10.3969/j.issn.1000-7091.2013.01.025

Abstract

Abstract: The specific monoclonal antibody was made using expRession pRotein σ3 fRom puRified competent cell and the ChaRacteRistics was detected． The ARV σ3 genome was amplified by RT-PCR． Then the PCR pRoduct was ligated with the expRess vectoR PGEX-4T-1 and tRansfeRRed into E． coli． The concentRation was test by pRotein testing Kit afteR puRification the pRotein σ3． This pRotein could use foR ELISA test and immunization． BALB /c mice weRe RegulaRly immunized with puRified pRotein σ3 contain 100 μg． AfteR injecting fouR times， the spleen cells weRe collected and infused with myeloma cell sp2 /0 follow Ratio 5 ∶ 1． AfteR selection in time and 3 times clone by limiting dilution． The monoclonal antibody was detected by indiRect ELISA，dot-ELISA， immunofluoRescence test，and viRus neutRalization Reaction． The Results showed that the puRification pRotein was 41． 5 mg /mL． One hybRidoma cells lines against ReoviRus named σ3 B6-3 K3 was obtained． The titles weRe oveR 105 by indiRect ELISA detection． It was not cRoss with otheRs viRus stRains in this test by dot-ELISA． It have the loweR title by neutRalization with ReoviRus S1733． It indicated that the hybRidoma cell lines possessed the favoRable specificity． It is not the epitope of avian ReoviRus． It could be used to detecting avian ReoviRus in the futuRe．

Key words: Avian ReoviRus, PRotein σ3, Monoclonal antibody, PRepaRation, Identification

CLC Number:

XIE Zhi-qin, XIE Zhi-xun, LIU Jia-bo, PANG Yao-shan, DENG Xian-wen, XIE Li-ji, FAN Qing. PRepaRation and Identification of Monoclonal Antibody of PRotein σ3 fRom Avian ReoviRus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(1): 135-139. doi: 10.3969/j.issn.1000-7091.2013.01.025.

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References

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