Partial purification of type A and B flagellin from Pseudomonas aeruginosa for preclinical use

Mohammed Jaafar  Al-Anssari; Alaa H. Al-Charrakh

doi:10.53730/ijhs.v6nS1.5590

Authors

Mohammed Jaafar Al-Anssari Dept. of Microbiology/ College of Medicine / University of Babylon, Hilla, Iraq and College of Medical and Health Technologies / University of AL-Kafeel, Al-Najaf, Iraq
Alaa H. Al-Charrakh
ahani67@gmail.com
Dept. of Microbiology/ College of Medicine / University of Babylon, Hilla, Iraq

Keywords:

Pseudomonas aeruginosa, Flagellin, Purification, Endotoxin Removal, Endotoxin detection, fliC gene

Abstract

Background: Due to their importance in vaccination, we tried in this study to purify the flagellin type a and type b separately from P. aeruginosa by a simple and cheap method with acceptable purity for preclinical use. Methods: Specific primer was used to detect the type a and b flagellin gene (fliC gene) in P. aeruginosa clinical isolates. Flagellin was purified from P. aeruginosa according to standard procedure with some modifications. Protein concentration and protein absorption were measured using a special spectrophotometer at 270 wavelength. Endotoxin removal was done using a special column. Then, the presence of flagellin was detected by SDS-PAGE. Results: The results showed two types of flagllin genes in different strains; type a (1020 bp) and type b (1250 pb) that amplified by PCR. Results also showed an appropriate concentration of purified protein by the used method with acceptable purity and decreasing the endotoxin to an acceptable level for preclinical use (less than 2.2 EU/ml). Conclusion: This research focused on the isolation and purification of flagellin protein from P. aeruginosa.

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