Inhibitory Monoclonal Antibodies and Their Recombinant Derivatives Targeting Surface-Exposed Carbonic Anhydrase XII on Cancer Cells

Autor: Lina Baranauskiene, Aurelija Zvirbliene, Vilma Petrikaite, Aiste Sliziene, Dovile Stravinskiene
Rok vydání: 2020
Předmět:
0301 basic medicine
single-chain fragment variable
law.invention
lcsh:Chemistry
0302 clinical medicine
law
Cricetinae
recombinant antibody
lcsh:QH301-705.5
Spectroscopy
Cells
Cultured
Carbonic Anhydrases
biology
Chemistry
Chinese hamster ovary cell
General Medicine
Recombinant Proteins
Computer Science Applications
030220 oncology & carcinogenesis
Monoclonal
Recombinant DNA
Antibody
circulatory and respiratory physiology
medicine.drug_class
carbonic anhydrase XII
cancer immunotherapy
monoclonal antibody
hybridoma
CHO Cells
Monoclonal antibody
Catalysis
Article
Inorganic Chemistry
03 medical and health sciences
Cricetulus
medicine
Animals
Humans
Physical and Theoretical Chemistry
Molecular Biology
Organic Chemistry
Molecular biology
030104 developmental biology
Epitope mapping
HEK293 Cells
lcsh:Biology (General)
lcsh:QD1-999
Cell culture
A549 Cells
Cancer cell
biology.protein
Epitope Mapping
Single-Chain Antibodies
Zdroj: International Journal of Molecular Sciences
International Journal of Molecular Sciences, Vol 21, Iss 9411, p 9411 (2020)
International Journal of Molecular Sciences, Basel : MDPI, 2020, vol. 21, no. 24, 9411, p. 1-26
International journal of molecular sciences, Basel : MDPI, 2020, vol. 21, no. 24, art. no. 9411, p. 1-26
Volume 21
Issue 24
ISSN: 1422-0067
1661-6596
Popis: Monoclonal and recombinant antibodies are widely used for the diagnostics and therapy of cancer. They are generated to interact with cell surface proteins which are usually involved in the development and progression of cancer. Carbonic anhydrase XII (CA XII) contributes to the survival of tumors under hypoxic conditions thus is considered a candidate target for antibody-based therapy. In this study, we have generated a novel collection of monoclonal antibodies (MAbs) against the recombinant extracellular domain of CA XII produced in HEK-293 cells. Eighteen out of 24 MAbs were reactive with cellular CA XII on the surface of live kidney and lung cancer cells as determined by flow cytometry. One MAb 14D6 also inhibited the enzymatic activity of recombinant CA XII as measured by the stopped-flow assay. MAb 14D6 showed the migrastatic effect on human lung carcinoma A549 and renal carcinoma A498 cell lines in a &lsquo
wound healing&rsquo
assay. It did not reduce the growth of multicellular lung and renal cancer spheroids but reduced the cell viability by the ATP Bioluminescence assay. Epitope mapping revealed the surface-exposed amino acid sequence (35-FGPDGENS-42) close to the catalytic center of CA XII recognized by the MAb 14D6. The variable regions of the heavy and light chains of MAb 14D6 were sequenced and their complementarity-determining regions were defined. The obtained variable sequences were used to generate recombinant antibodies in two formats: single-chain fragment variable (scFv) expressed in E. coli and scFv fused to human IgG1 Fc fragment (scFv-Fc) expressed in Chinese Hamster Ovary (CHO) cells. Both recombinant antibodies maintained the same specificity for CA XII as the parental MAb 14D6. The novel antibodies may represent promising tools for CA XII-related cancer research and immunotherapy.
Databáze: OpenAIRE
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