Government Degree College, Prithvipur District, Niwari, 472336, India.
Biotechnology Division, Defence Research & Development Establishment, Gwalior, 474 002, India.
Biotechnol Lett. 2021 May;43(5):1019-1036. doi: 10.1007/s10529-021-03091-4. Epub 2021 Feb 25.
To identify immunogenic proteins of C. botulinum type B secretome by immunoproteomic analysis.
In the present study, an attempt was made to elucidate the vaccine candidates/diagnostic molecules against botulism using immuno proteomic approach. C. botulinum type B secretome was elucidated when it was grown in TPGY as well as CMM media. Predominant 51 proteins were identified in both the media using 2-DE and mass spectrometry analysis. 2D gels (CMM & TPGY) were probed with respected proteins mice antiserum and obtained 17 and 10 immunogenic proteins in TPGY as well as CMM media respectively. Hypothetical protein CLOSPO_00563, ornithine carbamoyl transferase, FlaA, molecular chaperone GroEL and secreted protease proteins were found as the common immuno dominant proteins in both media. Polyclonal Antibodies raised against C. botulinum types A and E showed cross-reactivity with secretome C. botulinum type B at the lowest dilution (1:1000) but did not show cross reactivity with highest dilution (1:30,000) with C. botulinum type B secretome. Polyclonal antibodies against C. botulinum type F secretome did not show cross reactivity with C. botulinum type B secretome.
Identified immunogenic proteins can be used as vaccine candidates and diagnostic markers for the infant and wound botulism but common immunogenic proteins may be the best vaccine candidate molecule for development of vaccine as well as diagnostic system against the infant and wound botulism.
通过免疫蛋白质组学分析鉴定 B 型肉毒梭菌分泌组中的免疫原性蛋白。
本研究尝试使用免疫蛋白质组学方法阐明针对肉毒中毒的疫苗候选物/诊断分子。当 B 型肉毒梭菌在 TPGY 培养基和 CMM 培养基中生长时,阐明了其分泌组。使用 2-DE 和质谱分析在两种培养基中均鉴定出 51 种主要蛋白质。用相应的蛋白质小鼠抗血清探测 2D 凝胶(CMM 和 TPGY),在 TPGY 培养基和 CMM 培养基中分别获得 17 种和 10 种免疫原性蛋白。假设蛋白 CLOSPO_00563、鸟氨酸氨甲酰转移酶、FlaA、分子伴侣 GroEL 和分泌蛋白酶蛋白被发现是两种培养基中共同的免疫显性蛋白。针对 A 型和 E 型肉毒梭菌的多克隆抗体在最低稀释度(1:1000)与 B 型肉毒梭菌分泌组发生交叉反应,但在最高稀释度(1:30000)时与 B 型肉毒梭菌分泌组不发生交叉反应。针对 F 型肉毒梭菌分泌组的多克隆抗体与 B 型肉毒梭菌分泌组不发生交叉反应。
鉴定出的免疫原性蛋白可作为婴儿和创伤性肉毒中毒的疫苗候选物和诊断标志物,但共同的免疫原性蛋白可能是开发婴儿和创伤性肉毒中毒疫苗和诊断系统的最佳疫苗候选物分子。