The Effect of Differentially Designed Fusion Proteins to Elicit Efficient Anti-human Thyroid Stimulating Hormone Immune Responses

The Effect of Differentially Designed Fusion Proteins to Elicit Efficient Anti-human Thyroid Stimulating Hormone Immune Responses

The production of human thyroid stimulating hormone (hTSH) immunoassays requires specific antibodies against hTSH which is a cumbersome process. Therefore, producing specific polyclonal antibodies against engineered recombinant fusion hTSH antigens would be of great significance. The best immunogeni...

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Bibliographic Details
Published in:Iranian journal of allergy, asthma, and immunology Vol. 17; no. 2; pp. 158 - 170
Main Authors: Mard-Soltani, Maysam, Rasaee, Mohamad Javad, Khalili, Saeed, Sheikhi, Abdol-Karim, Hedayati, Mehdi, Ghaderi-Zefrehi, Hossein, Alasvand, Milad
Format: Journal Article
Language:English
Published: Iran Tehran University of Medical Sciences 01-04-2018
Subjects:
Antigens
Cross reactivity
Epitopes
Fusion protein immunization
Immune response
Immunization
Immunoassay
Immunoassays
Immunogenicity
Polyclonal antibodies
Protein folding
Protein purification
Proteins
Serum
Tetanus
Tetanus toxin
Thyroid
Thyroid gland
Titration
Immunoassay
Fusion protein immunization
Cross reactivity
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Summary:The production of human thyroid stimulating hormone (hTSH) immunoassays requires specific antibodies against hTSH which is a cumbersome process. Therefore, producing specific polyclonal antibodies against engineered recombinant fusion hTSH antigens would be of great significance. The best immunogenic region of the hTSH was selected based on in silico analyses and equipped with two different fusions. Standard methods were used for protein expression, purification, verification, structural evaluation, and immunizations of the white New Zealand rabbits. Ultimately, immunized serums were used for antibody titration, purification and characterization (specificity, sensitivity and cross reactivity). The desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. Structural analyses indicated that only the bigger antigen has showed changed 2 dimensional (2D) and 3D structural properties in comparison to the smaller antigen. The raised polyclonal antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum and negligible. The fusion which was solely composed of the tetanus toxin epitopes led to better protein folding and was capable of immunizing the host animals resulting into high titer antibody. Therefore, the minimal fusion sequences seem to be more effective in eliciting specific antibody responses.
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ISSN:1735-1502
1735-5249