Production and Characterization of a Monoclonal Antibody Against Spike Protein of Transmissible Gastroenteritis Virus

Abstract

Transmissible gastroenteritis virus (TGEV) is a member of coronaviruses. The viral spike (S) protein mediates the interaction between TGEV and its susceptible cells. Here, we expressed a truncated gene encoding the N terminal half of TGEV S gene (designated S1 gene) in a prokaryotic system. The resulting S1 protein was used to immunize BALB/c mice followed by the generation of a monoclonal antibody (MAb). A generated MAb (7F9) was identified by ELISA and the chromosome number of the hybridoma cell was analyzed. The immunoreactivity of the MAb to TGEV S protein was confirmed by Western blot analysis. Moreover, immunofluorescence assays showed that the MAb is able to detect cell infection by TGEV. The MAb achieved in this study can be used as a specific diagnostic reagent for detecting TGEV S protein.

Generation and Characterization of Antibody Against Porcine Interleukin-18

“…The E. coli system has advantages such as low cost, high production, and manipulation convenience, (14) and recently, we have expressed several heterologous proteins in this system successfully. (15)(16)(17)(18)(19) The SDS-PAGE showed that pIL-18 protein is approximately 25 kDa and the pIL-18 accumulated in the bacteria as inclusion bodies (Fig. 2).…”

“…Previously, we have purified several inclusion body proteins from bacteria using the gel-purification method. (15)(16)(17)(18)(19) However, the purified protein was denatured and subsequent renaturation is required in order to get a biologically active protein. In this study, we utilized the fused Histidine (His)-tag in the vector to purify the pIL-18 using a Ni-NTA affinity column.…”

A Novel Monoclonal Antibody Against the Constant Region of Goose Immunoglobulin Light Chain

“…The chromosome number of hybridoma cells was analyzed by the conventional method using colchicines (Sigma-Aldrich) with the control of SP2/0 cells. (13,14) The stability of passage and cryopreservation was also measured by conventional techniques. (13,15) The ascites of MAb against GoIgCL was prepared using standard procedures as described previously (16) and purified by affinity chromatography using Protein G (Genscript).…”

Preparation and Characterization of Polyclonal Antibody Against Severe Acute Respiratory Syndrome-associated Coronavirus Spike Protein

“…Our previous and current results confirm that the bacterially expressed heterologous proteins are functional immunogens for generating both monoclonal and polyclonal antibodies. (22)(23)(24)(25)28) Biological activity of the SARS-CoV S1…”