Related Experiment Video
Updated: Mar 29, 2026
Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
Published on: March 24, 2017
[Preparation and application of monoclonal antibody against human LOC339524 protein]
Zonghong Long1, Hong Li2, Feng Chen1
1Department of Anesthesiology, Second Affiliate Hospital, Third Military Medical University, Chongqing 400037, China.
Objective:
To prepare and apply monoclonal antibody (mAb) against human LOC339524 protein.
Methods:
The non-glycosylated antigenic gene sequence of the LOC339524 protein was expressed in triplicate to enhance immunogenicity. Then this synthetic gene was connected to pET28a plasmid. Recombinant LOC339524 protein was obtained by E.coli expression system and was administered intraperitoneally as an immunogen to BALB/c mice to obtain mAb. The specificity and titer of the mAb were characterized by ELISA. Recombinant LOC339524 protein was identified through Western blotting. The expression of the LOC339524 protein in human myocardial tissues and H9C2 cells were detected by immunohistochemistry, and its level in the sera of patients with different heart diseases was detected with the antibody.
Results:
We obtained two hybridoma cell lines, 5-D3 and 4-F8, secreting specific mAbs against LOC339524 protein. The titer of 5-D3 was up to 2×10(6), higher than the titer of 4-F8. Western blotting and immunohistochemistry demonstrated that 5-D3 could specifically recognize LOC339524 protein of Homo sapiens and Rattus norvegicus. With the antibody we obtained, we successfully detected the serum level of LOC339524 protein in patients with different heart diseases.
Conclusion:
The mAb against human LOC339524 protein with good specificity and high titer has been successfully prepared.
