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Article

Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization

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Laboratory of Structural Biology, Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic
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The Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, 1550 Orleans St, Baltimore, MD 21231, USA
3
Mass Spectrometry Group, Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, IOCB & Gilead Research Center, Flemingovo náměstí 542/2, 160 00 Prague, Czech Republic
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(18), 6672; https://doi.org/10.3390/ijms21186672
Received: 18 August 2020 / Revised: 8 September 2020 / Accepted: 9 September 2020 / Published: 12 September 2020
Prostate-Specific Membrane Antigen (PSMA) is an established biomarker for the imaging and experimental therapy of prostate cancer (PCa), as it is strongly upregulated in high-grade primary, androgen-independent, and metastatic lesions. Here, we report on the development and functional characterization of recombinant single-chain Fv (scFv) and Fab fragments derived from the 5D3 PSMA-specific monoclonal antibody (mAb). These fragments were engineered, heterologously expressed in insect S2 cells, and purified to homogeneity with yields up to 20 mg/L. In vitro assays including ELISA, immunofluorescence and flow cytometry, revealed that the fragments retain the nanomolar affinity and single target specificity of the parent 5D3 antibody. Importantly, using a murine xenograft model of PCa, we verified the suitability of fluorescently labeled fragments for in vivo imaging of PSMA-positive tumors and compared their pharmacokinetics and tissue distribution to the parent mAb. Collectively, our data provide an experimental basis for the further development of 5D3 recombinant fragments for future clinical use. View Full-Text
Keywords: prostate-specific membrane antigen; in vivo imaging; prostate cancer; monoclonal antibody; antibody fragment; glutamate carboxypeptidase II; NAALADase prostate-specific membrane antigen; in vivo imaging; prostate cancer; monoclonal antibody; antibody fragment; glutamate carboxypeptidase II; NAALADase
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MDPI and ACS Style

Novakova, Z.; Belousova, N.; Foss, C.A.; Havlinova, B.; Gresova, M.; Das, G.; Lisok, A.; Prada, A.; Barinkova, M.; Hubalek, M.; Pomper, M.G.; Barinka, C. Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization. Int. J. Mol. Sci. 2020, 21, 6672. https://doi.org/10.3390/ijms21186672

AMA Style

Novakova Z, Belousova N, Foss CA, Havlinova B, Gresova M, Das G, Lisok A, Prada A, Barinkova M, Hubalek M, Pomper MG, Barinka C. Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization. International Journal of Molecular Sciences. 2020; 21(18):6672. https://doi.org/10.3390/ijms21186672

Chicago/Turabian Style

Novakova, Zora, Nikola Belousova, Catherine A. Foss, Barbora Havlinova, Marketa Gresova, Gargi Das, Ala Lisok, Adam Prada, Marketa Barinkova, Martin Hubalek, Martin G. Pomper, and Cyril Barinka. 2020. "Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization" International Journal of Molecular Sciences 21, no. 18: 6672. https://doi.org/10.3390/ijms21186672

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