Application of Magnetic Particles for Fast Determination of Immunoreactive Fraction of 68Ga-Labelled VHH Antibodies to PD-L1.

IF 1.1 Q4 MEDICINE, RESEARCH & EXPERIMENTAL
Sovremennye Tehnologii v Medicine Pub Date : 2023-01-01 Epub Date: 2023-05-28 DOI:10.17691/stm2023.15.3.03
K O Avrov, S V Shatik, V V Zaitsev, R I Al-Shehadat, O A Shashkova, L A Terekhina, I S Malakhov, M P Samoylovich
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引用次数: 0

Abstract

Quantification of the immunoreactive fraction (IRF) of radioactive isotope-labeled antibodies or their fragments is necessary to assess the specific activity of radiopharmaceuticals. Traditionally, cells expressing the target molecules on their surface are used to determine IRF, but such analysis is time-consuming and has difficulties with standardization. The aim of the study was to develop a fast and reliable method for quantitative determination of IRF by 68Ga-labeled VHH antibodies to PD-L1 based on the use of magnetic particles coated with antigen molecules.

Materials and methods: Commercially available magnetic particles coated with protein A have been used in our study. The antigen conjugated with the Fc fragment (PD-L1-Fc) was immobilized on the particles. The IRF value of 68Ga radionuclide-labeled nanobodies (VHH) against PD-L1 (68Ga-VHH-PD-L1) was determined using magnetic particles coated with antigen molecules and cells expressing the antigen on their surface. When VHH antibodies were conjugated to 68Ga radionuclide, protein molecules were modified using bifunctional chelating agents: tetraazacyclododecanetetraacetic acid (DOTA) or deferoxamine (DFO). The magnitude of IRF was defined as the ratio of radioactivity specifically bound to particles or cells to the total radioactivity added to the sample.

Results: The specificity of the 68Ga-VHH-PD-L1 radioimmunoconjugate binding to the antigen-coated magnetic particles has been proved. Some special aspects, which should be taken into consideration when using this method, have been established. The comparison of the IRF estimates using the antigen-expressing cells and magnetic particles has not revealed any significant differences in the results obtained in our study. Nevertheless, the presented method based on magnetic particles with immobilized antigen molecules requires only 15 min to determine the radioimmunoconjugate IRF, which is of fundamental importance for the routine assessment of the specificity of radiopharmaceuticals containing short-lived isotopes.

磁颗粒法快速测定68ga标记的VHH PD-L1抗体免疫反应部分
放射性同位素标记的抗体或其片段的免疫反应部分(IRF)的定量对于评估放射性药物的特异性活性是必要的。传统上,在其表面表达靶分子的细胞用于测定IRF,但这种分析耗时且难以标准化。本研究的目的是开发一种快速可靠的方法,通过使用抗原分子包被的磁性颗粒,用68Ga标记的PD-L1 VHH抗体定量测定IRF
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来源期刊
Sovremennye Tehnologii v Medicine
Sovremennye Tehnologii v Medicine MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
1.80
自引率
0.00%
发文量
38
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