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368466

An optimized protocol for the expression of a recombinant protein fragment of human factor B used for the production of polyclonal antibodies

Article

Last updated: 28 Dec 2024

OverviewSimilar Items

Subjects

-

Tags

chemotherapy

Abstract

Background: Complement is a critical component of the innate immunity bridging both arms of the immune system. Extensive trials are ongoing to develop therapeutics that target complement and hence alleviating complement-mediated disorders. Objectives: Here, we detail our attempts to express a recombinant protein fragment of human factor B (FBfr) in a high yield and solubility in addition to the production of polyclonal antibody against it. Methodology: We tested different expression conditions and host strains to express FBfr. Moreover, we used the expressed Fbfr to immunize rabbits for production of polyclonal antibody and tested its inhibitory properties against the alternative pathway by in vitro assays. Results: We successfully expressed an alternative complement pathway inhibitor FBfr in E. coli using an optimized protocol. The protein was antigenic and led to the production of a high titer of antibodies in sera of the immunized rabbits. The polyclonal antibody was shown to have a moderate inhibition of the alternative complement pathway activity. Conclusion: optimization of expression parameters can greatly improve the expression levels of recombinant proteins. These proteins can have a functional activity and be used for successful immunization process.

DOI

10.21608/ejmm.2024.301876.1272

Keywords

alternative pathway, complement, factor B, factor B fragment, Inhibitor

Authors

First Name

Enas

Last Name

Sultan

MiddleName

Y.

Affiliation

Department of Microbiology & Immunology, Faculty of Pharmacy, Mansoura University; Egypt

Email

dr_noosa_2009@yahoo.com

City

Mansoura

Orcid

-

First Name

Dina

Last Name

Rizk

MiddleName

E.

Affiliation

Department of Microbiology & Immunology, Faculty of Pharmacy, Mansoura University; Egypt

Email

dinaeid2013@yahoo.com

City

-

Orcid

-

First Name

Ramadan

Last Name

Hassan

MiddleName

-

Affiliation

Department of Microbiology & Immunology, Faculty of Pharmacy, Mansoura University; Egypt

Email

rhassan33@gmail.com

City

-

Orcid

-

First Name

Hany

Last Name

Kenawy

MiddleName

I.

Affiliation

Department of Microbiology & Immunology, Faculty of Pharmacy, Mansoura University; Egypt

Email

hanykenawy@mans.edu.eg

City

Mansoura

Orcid

0000-0002-8548-6649

Volume

33

Article Issue

3

Related Issue

47674

Issue Date

2024-07-01

Receive Date

2024-07-05

Publish Date

2024-07-01

Page Start

141

Page End

151

Print ISSN

1110-2179

Online ISSN

2537-0979

Article File

EJMM_Volume 33_Issue 3_Pages 141-151.pdf

PDF . 781.3kB

Link

https://ejmm.journals.ekb.eg/article_368466.html

Detail API

https://ejmm.journals.ekb.eg/service?article_code=368466

Order

368,466

Type

New and original researches in the field of Microbiology.

Type Code

2,038

Publication Type

Journal

Publication Title

Egyptian Journal of Medical Microbiology

Publication Link

https://ejmm.journals.ekb.eg/

MainTitle

An optimized protocol for the expression of a recombinant protein fragment of human factor B used for the production of polyclonal antibodies

Details

Type

Article

Created At

28 Dec 2024