Professor Peng Dapeng of Huazhong Agricultural University: ic-ELISA detection method and molecular recognition mechanism based on kitatamycin monoclonal antibody in animal tissues

Original title: Professor Peng Dapeng of Huazhong Agricultural University: ic-ELISA detection method and molecular recognition mechanism based on kitatamycin monoclonal antibody in animal tissues

Recently, the team of Professor Peng Dapeng of Huazhong Agricultural University prepared KIT-specific antigens and developed monoclonal antibodies, and established ic-ELISA methods and sample preparation methods in animal tissues to monitor KIT residues.

Abstract

In order to monitor the residue of kitasamycin (KIT), an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on the KIT-specific monoclonal antibody KA/2A9 was established.

Introduction

Kitasamycin (KIT) is a macrolide antibiotic with a 16-membered ring (Figure S1) (Arsic, Barber, Čikoš, Mladenovic, Stankovic, & Novak, 2018).

Materials and methods

1.

Single chain antibody phage library preparation

The collected KA/2A9 hybridoma cells (1×107) were centrifuged at 1300 r/min for 10 min at room temperature.

Panning and soluble expression of KIT-specific phage single-chain antibody

Using M13K07 helper phage, the constructed phage library was used to prepare phage antibodies.

Single-chain antibody sequence analysis and three-dimensional structure prediction

The ScFv nucleotide sequence was sequenced by Shenggong Biotech (Shanghai, China).

Results and Discussion

1.

According to reports, carboxymethoxy amine hemihydrochloride (CMO) is often used as a spacer to design haptens, and is used to develop specific monoclonal antibodies for erythromycin, tylosin and other small molecule drugs ( Lai et al.

2.

The monoclonal hybridoma cell lines KA/1H8 and KA/2A9 were selected for the preparation of mouse ascites.

3.

Figure 1.

4.