Production and Characterization of Monoclonal Antibodies against a Recombinant Nfa1 Protein Cloned from Pathogenic Naegleria fowleri

OBJECTIVE: In order to use in the immunopathological studies of pathogenic Naegleria fowleri, monoclonal antibodies against a recombinant His-tag fusion Nfa1 protein cloned from pathogenic N. fowleri were produced, and its characterization was elucidated. In addition, to observe the usage in immunological studies, the localization of the Nfa1 protein in N. fowleri was observed by immunofluorescent assay using monoclonal antibodies. METHODS: Six-week-old female BALB/c mice were immunized with a purified recombinant His-tag fusion Nfa1 protein. Cell lines producing McAbs against the recombinant His-tag fusion Nfa1 protein were produced according to the protocol of hybridoma technique using myeloma cells (F0 cell). The McAbs were characterized by isotyping and Western blot analysis. RESULTS: Eight McAbs (A1F, A3C, A6D, A10F, C3A, C6F, C7F, B6B) against the rNfa1 protein were obtained. Eight McAbs isotypes were all IgG2b. The McAbs had high affinity against the recombinant His-tag fusion Nfa1 protein, which was resulted by indirect ELISA. They recognize 17 kDa protein which was defined with the recombinant His-tag fusion Nfa1 protein by Western blot analysis. Also, The Nfa1 protein reacted with eight McAbs showed on pseudopodia and food-cups by a fluorescent microscope. CONCLUSION: Eight hybridoma cell lines (A1F, A3C, A6D, A10F, C3A, C6F, C7F, B6B) secreting high titer of McAbs against the recombinant His-tag fusion Nfa1 protein were established. Also, the localization of the Nfa1 protein reacted with eight McAbs was observed on pseudopodia and food-cups of N. fowleri trophozoites by a fluorescent microscope. Therefore, the anti-Nfa1 monoclonal McAbs will be efficiently used in immunological study of PAME by N. fowleri.

Author dc:creator

• 이, 양진

Contributors dc:contributor

• 신, 호준
• 대학원 의학과
• 105424

• 파울러자유아메바
• Naegleria fowleri
• 원발성 아메바성 수막뇌염
• 유전자 클로닝
• 재조합 His-tag fusion

Language dc:language

ko

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000000617
000000000617

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/1494