Bioinformatic prediction and experimental validation of a PE38-based recombinant immunotoxin targeting the Fn14 receptor in cancer cells.

Abstract

AIM:

AFn14R can serve as an ideal target for cancer immunotherapy. Here, a combined bioinformatic and experimental approach was applied to characterize an immunotoxin consisting of single-chain variable fragment antibody that targets Fn14 and a toxin fragment (PE38).

METHODS & RESULTS:

Flow cytometry results showed that the rate of PE38-P4A8 binding to Fn14 was approximately 60 and 40% in HT-29 and A549 cells, respectively. Moreover, 1 ng/µl of immunotoxin was able to lyse approximately 53 and 41% of HT-29 and A549, respectively. PE38-P4A8 showed stability in mouse serum (∼90%) after 3-h incubation. Most importantly, using bioinformatics for determining the structure and function of fusion proteins can be very helpful in designing of experiments.

CONCLUSION:

Coupled with bioinformatics, experimental approaches revealed that PE38-P4A8 could be used as a promising therapeutic agent for cancer cells expressing Fn14.

• Authors: Maryam Keshtvarz, Jafar Salimian, Mehdi Yaseri, Seyedeh Zahra Bathaie, Ehsan Rezaie, Amir Aliramezani, Zahra Norouzbabaei, Jafar Amani, Masoumeh Douraghi
• Journal: Immunotherapy (Immunotherapy) Vol. 9 Issue 5 Pg. 387-400 (Mar 2017) ISSN: 1750-7448 [Electronic] England
• PMID: 28357912 (Publication Type: Journal Article, Validation Study)

Chemical References

• Antigens, Neoplasm
• Immunotoxins
• Receptors, Tumor Necrosis Factor
• Recombinant Fusion Proteins
• Single-Chain Antibodies
• TNFRSF12A protein, human
• TWEAK Receptor
• Tnfrsf12a protein, mouse

Topics

• A549 Cells
• Adenocarcinoma (therapy)
• Antigens, Neoplasm (metabolism)
• Computational Biology
• Humans
• Immunotoxins (genetics, metabolism)
• Molecular Targeted Therapy
• Receptors, Tumor Necrosis Factor (metabolism)
• Recombinant Fusion Proteins (genetics)
• Single-Chain Antibodies (genetics, metabolism)
• TWEAK Receptor