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Journal code(s)
- E-ISSN Number: 2277-7695
- P-ISSN Number: 2349-8242
- CODEN Code: PIHNBQ
- ZDB-Number: 2663038-2
- IC Journal ID: 7725
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Vol. 10, Special Issue 11 (2021)
In vitro effect of single-domain antibody, selected from the phage display library of Indian desert camel, on Pasteurella multocida LPS-induced activation of bovine peripheral leukocytes
Author(s):
Akhil Kumar Gupta, Anu Malik, Ajit Singh, Naresh Kumar Kakker and Jagveer Rawat
Akhil Kumar Gupta, Anu Malik, Ajit Singh, Naresh Kumar Kakker and Jagveer Rawat
Abstract:
The objective of the present study was to assess the ability of single-domain antibody clone 26 (dAb Cl 26) selected from phage display library to reduce Pasteurella multocida (vaccine strain p52) LPS -induced effects on buffalo leukocytes in vitro. The dAb Cl26 was expressed to an optimum level and later purified under denaturing condition using Ni-NTA chromatography. The protein concentration of purified dAb Cl-26 was estimated by BCA method and found to be 280 µg/ml. LPS of vaccine strain (p52) of P. multocida was extracted and concentration was found to be 38 µg/ml. The purity of extracted LPS was assessed for nucleic acid and protein contamination and was found to be negative. The purified dAb Cl-26 showed reactivity with Pasteurella multocida (p52) LPS by Indirect ELISA as well as Dot-Immunoblotting.
The objective of the present study was to assess the ability of single-domain antibody clone 26 (dAb Cl 26) selected from phage display library to reduce Pasteurella multocida (vaccine strain p52) LPS -induced effects on buffalo leukocytes in vitro. The dAb Cl26 was expressed to an optimum level and later purified under denaturing condition using Ni-NTA chromatography. The protein concentration of purified dAb Cl-26 was estimated by BCA method and found to be 280 µg/ml. LPS of vaccine strain (p52) of P. multocida was extracted and concentration was found to be 38 µg/ml. The purity of extracted LPS was assessed for nucleic acid and protein contamination and was found to be negative. The purified dAb Cl-26 showed reactivity with Pasteurella multocida (p52) LPS by Indirect ELISA as well as Dot-Immunoblotting.
For in vitro testing of purified dAb Cl-26 on P. multocida (p52) LPS-induced activation of bovine peripheral leukocytes, Buffalo peripheral blood plastic adherent mononuclear cells were exposed to LPS alone and in combination with different concentration of dAb Cl-26 and the expression level of various genes namely TNFα, IL-1β and IL-6 were studied by qRT-PCR. The dAb Cl-26, in a dose dependent manner, reduced the LPS-induced TNFα, IL-1β and IL-6 transcripts in buffalo peripheral leukocytes in vitro.
Pages: 2346-2351 | 324 Views 84 Downloads
How to cite this article:
Akhil Kumar Gupta, Anu Malik, Ajit Singh, Naresh Kumar Kakker and Jagveer Rawat. In vitro effect of single-domain antibody, selected from the phage display library of Indian desert camel, on Pasteurella multocida LPS-induced activation of bovine peripheral leukocytes. The Pharma Innovation Journal. 2021; 10(11S): 2346-2351.
Journal code(s)
- E-ISSN Number: 2277-7695
- P-ISSN Number: 2349-8242
- CODEN Code: PIHNBQ
- ZDB-Number: 2663038-2
- IC Journal ID: 7725
Main Menu
Special Issue
Copyright Form
Identifier
The Pharma Innovation Journal
