We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
The Effect of Differentially Designed Fusion Proteins to Elicit Efficient Anti-human Thyroid Stimulating Hormone Immune Responses.
- Authors
Mard-Soltani, Maysam; Rasaee, Mohamad Javad; Khalili, Saeed; Sheikhi, Abdol-Karim; Hedayati, Mehdi; Ghaderi-Zefrehi, Hossein; Alasvand, Milad
- Abstract
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.
- Subjects
CHIMERIC proteins; THYROTROPIN; IMMUNE response; PROTEIN expression; GLYCOPROTEIN hormones; AMINO acids; ANIMALS; ANTIGEN-antibody reactions; ANTIGENS; GENETIC techniques; IMMUNITY; IMMUNIZATION; IMMUNOGLOBULINS; MOLECULAR structure; NUCLEOTIDES; RABBITS; RECOMBINANT proteins; TETANUS toxin
- Publication
Iranian Journal of Allergy, Asthma & Immunology, 2018, Vol 17, Issue 2, p158
- ISSN
1735-1502
- Publication type
journal article