Porcine Anti-TGEV Recombinant Antibody (clone TZZ19)

Forty-eight clones were randomly selected from the library and screened, and three clones (TZZ14, TZZ 19 and TZZ43) with relatively high binding affinity were identified.

Zhang, F., Chen, Y., Yang, L., & Zhu, J. (2019). Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro. Archives of virology, 164(4), 983-994.

Lane 1, purified scFv TZZ 14; lane 2, purified scFv TZZ 19; lane 3, purified scFv TZZ 43; lane 4, E. coli BL21(DE3) transformed with pET-28a (+)

Zhang, F., Chen, Y., Yang, L., & Zhu, J. (2019). Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro. Archives of virology, 164(4), 983-994.

Diluted scFv antibody was incubated with TGEV; this was followed by infection of ST cells. The scFv antibodies showed no affinity for TGEV and were used as a negative control.

Zhang, F., Chen, Y., Yang, L., & Zhu, J. (2019). Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro. Archives of virology, 164(4), 983-994.

scFv TZZ 14, TZZ 19 and TZZ 43 inhibited TGEV infection. An unrelated scFv ZW 88 and TGEV-negative porcine serum were used as a negative control.

Zhang, F., Chen, Y., Yang, L., & Zhu, J. (2019). Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro. Archives of virology, 164(4), 983-994.

ST cells were infected with TGEV SHXB strains and incubated with purified scFv or anti-TGEV spike (S) protein monoclonal antibody followed by FITC-conjugated secondary antibody. The nuclei were counterstained with 4', 6'-diamidino-2-phenylindole (DAPI). Cells that were not infected with TGEV were used as a negative control and examined under a fluorescence microscope.

Zhang, F., Chen, Y., Yang, L., & Zhu, J. (2019). Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro. Archives of virology, 164(4), 983-994.

293T cells were transfected with recombinant plasmid pCDNA3.1(+)-S and incubated with scFv antibody, an unrelated scFv, ZW 88, or anti-TGEV spike (S) protein monoclonal antibody, followed by staining with FITC- conjugated secondary antibody. The cells were examined under a fluorescence microscope. Cells transfected with p3 × FLAG-CMV-14 followed by staining with mouse anti-TGEV S monoclonal antibody were used as an additional negative control.

Zhang, F., Chen, Y., Yang, L., & Zhu, J. (2019). Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro. Archives of virology, 164(4), 983-994.

The membrane containing the proteins was incubated with purified scFv, an unrelated scFv, ZW 88, or anti-TGEV spike (S) protein monoclonal antibody. Cells transfected with p3 × FLAG-CMV-14 followed by staining with mouse anti-TGEV S monoclonal antibody were used as an additional negative control

Zhang, F., Chen, Y., Yang, L., & Zhu, J. (2019). Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro. Archives of virology, 164(4), 983-994.

Three scFv antibodies were evaluated for cross-reactivity against the following swine pathogens: transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), porcine respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), ETEC enterotoxigenic Escherichia coli (ETEC) K88, and ETEC K99. BSA was used as a negative control.

Zhang, F., Chen, Y., Yang, L., & Zhu, J. (2019). Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro. Archives of virology, 164(4), 983-994.