Delving into the Latest Updates on TAT-HuScFv with Synapse

Author: ARREDONDO-ESPINOZA, EDER ; Arredondo-Espinoza, Eder ; GONZÁLEZ-CRUZ, ALDO O. ; González-Cruz, Aldo O ; Pioquinto-Ávila, Elizeth ; Zarate, Xristo ; BALDERAS-RENTERÍA, ISAÍAS ; ZARATE, XRISTO ; PIOQUINTO-ÁVILA, ELIZETH ; Balderas-Rentería, Isaías ; SOLIS-ESTRADA, JORGE ; Solis-Estrada, Jorge

BACKGROUND/AIM:

The epidermal growth factor receptor (EGFR) is over-expressed in several types of cancer, and monoclonal antibody therapy has been the strategy that has shown the best results. This study focused on the construction of a humanized single chain antibody (huscFv) directed against EGFR (HER1).

MATERIALS AND METHODS:

The CDR grafting method was used to incorporate murine complementarity determining regions (CDRs) of cetuximab into human sequences. A dot blot assay was used to examine the affinity of the huscFv secreted by HEK293T for EGFR. The inhibitory effect on the viability of A549 cells was evaluated using the WST-1 assay.

RESULTS:

The incorporation of murine CDRs of cetuximab into human sequences increased the degree of humanness by 16.4%. The increase in the humanization of scFv did not affect the affinity for EGFR. Metformin had a dose-dependent effect, with an IC₅₀ of 46 mM, and in combination with huscFv, the cell viability decreased by 45% compared to the 15% demonstrated by huscFv alone.

CONCLUSION:

The CDR grafting technique is efficient for the humanization of scFv, maintaining its affinity for EGFR and demonstrating its inhibitory effect when combined with metformin in A549 cells.

01 Jan 2024·ASIAN PACIFIC JOURNAL OF ALLERGY AND IMMUNOLOGY

Binding specificity of HuScFv to cholangiocarcinoma cells; New avenues for diagnosis and treatment

Article

Author: Chaicumpa, Wanpen ; Chulanetra, Monrat ; Duang Sa-Ard, Sunisa ; Sawatpiboon, Nathawadee ; Wongkajornsilp, Adisak ; Kasetsinsombat, Kanda

BACKGROUND:

Cholangiocarcinoma (CCA) is a very aggressive cancer of the bile ducts. Recent advances in immunotherapy, particularly with human single-chain variable fragments (HuScFv), have shown promise in the treatment of solid tumors by targeting cancer cells or improving the immune response.

OBJECTIVE:

This study aimed to select and produce human single-chain antibody fragments (HuScFv) specific to CCA cells (HubCCA1, RMCCA) from phage display HuScFv libraries with minimum or no binding to cholangiocytes (MMNK1).

METHODS:

Phages that displayed HuScFv to CCA cells were selected by bio-panning and each phagemid was transduced to HB2151 E. coli. The presence of huscfv was determined by direct colony PCR. HuScFv was produced in the E. coli and detected by Western blot analysis and confirmed their specificity and binding capacity to CCA by flow cytometry.

RESULTS:

From biopanning, 196 of 350 colonies (56%) of HB2151 E. coli harboring the huscfv gene, and 106 of these (30%) produced the protein. Flow cytometry testing with 14 clones confirmed the presence of the HuScFv protein. The result showed that five HuScFv clones (25, 33, 61, 68, and 80) exhibited stronger binding to CCA cell lines (HubCCA1, RMCCA) compared to cholangiocytes. Furthermore, each clone possessed a distinct amino acid sequence, suggesting unique binding specificities.

CONCLUSIONS:

HuScFv specific to CCA cells were successfully selected from phage display HuScFv libraries which offer new revenues to develop a pan-CCA immunotherapy and diagnosis of CCA.

100 Deals associated with TAT-HuScFv

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Indication	Highest Phase	Country/Location	Organization	Date
influenza A subtype H5N1 infection	Preclinical	CN	Jilin Agricultural University	31 Mar 2022
influenza A subtype H5N1 infection	Preclinical	CN	Chinese Academy of Agricultural Sciences	31 Mar 2022
influenza A subtype H5N1 infection	Preclinical	-	Changchun First Test of Chinese Patent Medicine Co., Ltd.	31 Mar 2022