Identification of a novel epitope specific for Gp85 protein of avian leukosis virus subgroup K

Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus

“…These results suggest that the antibody secretion of the prepared hybridoma cell line is superior to that described in previous reports. 25 …”

“…These results suggest that the antibody secretion of the prepared hybridoma cell line is superior to that described in previous reports. 25 It can be shown from the results in Figure 3B,C that the prepared MAb can react both with the expressed products of the pET32a-ALV-K plasmid (Figure 3B) and with the extracted gp85 protein from the ALV-K-JS11C1 strain (Figure 3C). In Figure 3C, the extracted viral proteins were digested with PNGase F enzyme (an amidohydrolase, which can cleave high mannose, heterozygous, and complex oligosaccharide from the glycoprotein linked by asparagine), and the digested products were analyzed by the western blot mediated with the prepared MAb; the results show a positive reaction at the electrophoretic band (39 kDa) of the digested protein.…”

“…The sequencing result shows that its sequence is completely consistent with that of the screened ALV-K-JS11C1 gene fragment in Table 2. Compared with the sequence obtained using the whole gp85 gene as a target gene in the previous report, 25 this strategy greatly improved the accuracy and efficiency of preparing a specific monoclonal antibody. The target gene was ligated into the pMD18-T vector to construct the recombinant amplification plasmid (pMD18T-ALV-K) and prepare enough target gene products.…”

“… 20 − 22 In our laboratory, an ALV-A monoclonal antibody was also successfully developed and utilized in a rapid and specific colloidal gold test strip. 23 , 24 Recently, a MAb against ALV-K that can recognize an epitope of the gp85 protein containing 17 amino acid residues ( 129– AFGPRSIDTLSDWSRPQ –145 ) has been reported; 25 however, it is difficult to apply it clinically due to the lack of reports about its biological characteristics such as specificity, stability, and antigenic epitope structure. In this study, a new MAb against another ALV-K gp85 epitope ( 217– RRNYT –221 ) was successfully prepared, and its biological characteristics were clarified, which contributes to the establishment of novel and specific detection methods for ALV-K.…”

Isolation and characterization of avian leukosis virus subgroup J associated with hemangioma and myelocytoma in layer chickens in China

“…The ALV env gene encodes envelope glycosylation protein, which is linked together with the surface glycoprotein subunit (SU) encoded by the gp85 gene, and the transmembrane glycoprotein subunit (TM) encoded by the gp37 gene into a rod-shaped dimer that is attached to the envelope surface, namely virus glycoprotein ( 9 ). Gp85 is the ligand of the virus utilized to invade host cells, which also serves as the foundation for subgroup categorization ( 10 ). The significance of long terminal repeat (LTR) and gp85 in the etiology of ALV-J has been confirmed by previous studies ( 11 ).…”