Reduced Tumor Volume and Increased Necrosis of Human Breast Tumor Xenograft in Mice Pretreated by a Cocktail of Three Specific Anti-HER2 scFvs


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Purpose:We aimed to assess the effects of a cocktail comprising three specific anti- HER2 scFvs on breast tumor formation in a xenograft mouse model and to evaluate quantitative changes in the tumor using stereological analysis.

Methods:Three specific anti-HER2 phage antibodies were produced from a scFv-library using phage display technology. The cell binding capacities of the antibodies were assessed via FACS analysis. Soluble forms of the antibodies were prepared by infecting HB2151-E. coli cells and purified using a centrifugal ultrafiltration method. The purification process was evaluated by SDSPAGE analysis. Two forms of scFv cocktails were prepared, soluble scFv and phage-scFv cocktail, which contained an equal amount/phage of each of the three antibodies. Inbred female BALB/c mice were pretreated with 5 and 20 mg/kg of the soluble scFv cocktail and 1011 phage-scFv cocktail/ kg. The mice were then injected with 2×106 SKBR-3 human breast cancer cells. Total tumor, inflammatory and non-inflammatory volumes were estimated using the Cavalieri principle after preparing photomicrograph slides.

Results:The anti-HER2 scFvs showed significantly higher binding to SKBR-3 cells compared to the isotype control. SDS-PAGE analysis confirmed the high purification of the scFvs. Stereological analysis revealed that the group pretreated with 20 mg/kg of the soluble scFv cocktail exhibited the highest reductions in total tumor volume, non-inflammatory volume, and inflammatory volume, with reductions of 73%, 78%, and 72%, respectively, compared to PBS-pretreated mice (P-valup < 0.0001). The volumetric ratio of necrotic tissue to total tumor volume increased by 2.2-fold and 2- fold in the 20mg/kg of soluble scFv cocktail and phage-scFv cocktail groups, respectively, compared to the PBS-treated mice (P-valup < 0.05).

Conclusion:Pre-treatment with a 20 mg/kg anti-HER2 scFv cocktail resulted in a significant reduction in tumor volume and increased necrotic area in a human breast cancer xenograft model, indicating the remarkable anti-tumor effect of the cocktail in vivo.

作者简介

Foroogh Nejatollahi

Recombinant Antibody Laboratory, Department of Immunology, Shiraz University of Medical Sciences

编辑信件的主要联系方式.
Email: info@benthamscience.net

Elham Nadimi

Recombinant Antibody Laboratory, Department of Immunology, Shiraz University of Medical Science

Email: info@benthamscience.net

Ali Noorafshan

Department of Neuroscience,, City University in Canada

Email: info@benthamscience.net

Setareh Moazen

Faculty of Pharmaceutical Sciences, University of British Columbia

Email: info@benthamscience.net

Ali Alizadeh

Cancer Research Center, Tehran University of Medical Sciences

Email: info@benthamscience.net

Solmaz Khalighfard

Cancer Research Center, Tehran University of Medical Sciences

Email: info@benthamscience.net

Amirhossein Sahebkar

Biotechnology Research Center, Pharmaceutical Technology Institute,, Institute, Mashhad University of Medical Sciences

Email: info@benthamscience.net

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