Characterization of the novel anti-TNF-<b>α</b> single-chain fragment antibodies using experimental and computational approaches

Pourtaghi-Anvarian, Samira; Mohammadi, Samaneh; Hamzeh‐Mivehroud, Maryam; Alizadeh, Ali; Dastmalchi, Siavoush

doi:10.1080/10826068.2018.1487855

Cited by 6 publications

(2 citation statements)

References 39 publications

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“…Cloning process was performed as described previously 34 . Briefly, D53 coding genes located in phage pR2 genome were amplified and inserted into pGEX-6P-1 vector.…”

Section: Cloning Of Dna Sequences Of D53 Into Pgex-6p-1mentioning

confidence: 99%

“…Briefly, D53 coding genes located in phage pR2 genome were amplified and inserted into pGEX-6P-1 vector. However, DNA sequencing revealed the presence of an amber stop codon in the CDR2 region of D53, which is a problem frequently observed in phage display libraries 34,35 . The problem was resolved using site directed point mutation using overlap extension PCR mutagenesis technique to convert TAG amber stop codon into CAG to code for amino acid glutamine.…”

Section: Cloning Of Dna Sequences Of D53 Into Pgex-6p-1mentioning

confidence: 99%

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Expression, purification and characterization of anti-FGF7 domain antibody identified using phage display technique

Alizadeh¹,

Roshani²,

Kandjani³

et al. 2021

Pharm Sci

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Background: Fibroblast growth factors (FGFs) are involved in angiogenesis, wound healing and embryonic development. However, one of the causes of cancer cell growth in fibroblast-dependent cancers is FGF7 secreted by fibroblasts. Therefore, antibodies against FGF7 can be used for treatment of these types of cancers. Methods: In previous studies, a phage displaying single domain antibody, D53, against human FGF7 has been identified using the phage display technique. In the present study, D53 was produced and purified in its isolated form. ELISA experiment was performed to evaluate the binding of D53 to FGF7. The mode of interaction of D53-FGF7 was explored using docking study and molecular dynamics (MD) simulations. Results: The expression and purification processes were verified using western blotting and SDS-PAGE analyses. ELISA experiment showed that D53 is able to recognize and bind FGF7. Docking study and MD simulations indicated that compared to dummy VH, D53 has more affinity towards FGF7. Conclusion: The findings in the current study can be useful for generation and development of FGF7 inhibitors with potential use in fibroblast-dependent cancers.

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“…Cloning process was performed as described previously 34 . Briefly, D53 coding genes located in phage pR2 genome were amplified and inserted into pGEX-6P-1 vector.…”

Section: Cloning Of Dna Sequences Of D53 Into Pgex-6p-1mentioning

confidence: 99%

Section: Cloning Of Dna Sequences Of D53 Into Pgex-6p-1mentioning

confidence: 99%