An improved method for analysis of technetium radiolabeled antibody by size exclusion HPLC

doi:10.1016/0969-8043(94)90169-4

Applied Radiation and Isotopes

Volume 45, Issue 10, October 1994, Pages 1009-1019

https://doi.org/10.1016/0969-8043(94)90169-4 Get rights and content

Abstract

A quantitative radio HPLC method for the analysis of radiolabeled ^99mTc compounds, in particular labeled antibodies or proteins, is described. The method is based on the quantitative re-oxidation of the reduced column absorbed ^99mTc species to pertechnetate using hydrogen peroxide. The pertechnetate is quantitatively eluted from the column. The method can be used as a stand alone technique for determining the true radiolabeling yield of labeled proteins/antibodies. The method has been automated for routine use and the typical analysis time is 30 min.

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Single radio UHPLC analysis for the quality control of technetium-99m radiolabelled radiopharmaceuticals
2021, Applied Radiation and Isotopes
Citation Excerpt :
Over a seven-month study in our radiopharmacy unit, more than 15% of the preparations had to be reanalysed using r-TLC, while only 0.7% of RP were considered as non-compliant. To increase method sensitivity and robustness, using other separative methods such as radio liquid chromatography can be a relevant option (Muddukrishna et al., 1994; Russell and Majerik, 1979). High performance liquid chromatography (HPLC) is preferentially employed for quality control by industrial companies.
The radiochemical purity (RCP) determination of radiopharmaceuticals is routinely done with radio-thin layer chromatography (r-TLC). These methods are usually transposed and adjusted from the summary product characteristics without any analytical validation. The r-TLC method is simple but manually-performed steps could lead to RCP misinterpretation. To increase the sensitivity, radio ultra-high performance liquid chromatography (r-UHPLC) can be used. In this study, an r-UHPLC method had been validated and compared to the r-TLC method. Hydrolyzed-reduced technetium had also been studied.
Technetium 1994
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Technetium speciation: Non-size effects in size-exclusion chromatography
1996, Journal of Chromatography A
The predominant separation technique used in technetium speciation, e.g., in plant material, is size-exclusion chromatography. However, interactions of technetium compounds with the stationary phase, i.e., non-size effects, have been reported. To evaluate these effects, the interaction of three technetium compounds, the pertechnetate anion (TcO₄⁻), the anionic Tc-diethylenetriaminepentaacetate complex (Tc-DTPA), and the cationic [Tc(V)O₂(1,4,8,11-tetraazacyclotetradecane)]⁺ complex (Tc-cyclam) with three different types of size-exclusion packing material, Sephadex G-25, Zorbax GF-250 and HEMA-SEC BIO 1000 has been studied. The effects of the composition of the mobile phase and the column temperature on the distribution coefficients of the three technetium compounds was measured. It is shown that non-size effects play a dominant role in the separation of these technetium compounds.
Direct <sup>99m</sup>Tc-labeling of antibodies by sodium dithionite reduction, and role of ascorbate as a stabilizer in cysteine challenge
1996, Nuclear Medicine and Biology
A method for the direct ^99mTc-labeling of antibodies by dithionite reduction was developed. Among three murine monoclonal IgG1 and one human polyclonal IgG (hIgG) antibodies tested, hIgG was the most quickly reduced by dithionite. These differences may reflect the reactivities of antibody disulfide bonds toward the oxidation products of dithionite. By optimizing reduction conditions to generate enough free sulfhydryl groups, it was possible to radiolabel human IgG and monoclonal antibody 170 with ^99mTc with a 90% monomeric antibody efficiency. The process avoided colloid formation. In contrast, about 0.1 sulfhydryl groups per antibody molecule, less than 1% of the possible 36, were detected after treatment with ascorbate (up to 35,000:1 molar ratio) at room temperature for 1 h for the antibodies tested. Sulfhydryl groups generated in antibodies were estimated using a new method: 5-iodoacetamidofluorescein-labeled antibodies quantitated by size exclusion HPLC. Ascorbate was found to prevent antibody aggregate formation in cysteine-challenged samples.
A caveat on the use of 2-mercaptoethanol in plant technetium speciation procedures
1996, Applied Radiation and Isotopes
2-mercaptoethanol, a reducing and chelating compound extensively used in plant technetium speciation procedures, is shown to react with the technetium-diethylenetriaminepentaacetate (Tc-DTPA) complex. The results of earlier plant technetium speciation studies should therefore be interpreted with caution, because of the possibility of reactions between labile plant technetium species and 2-mercaptoethanol.
Technetium Complexes - Their Possible Use in Radiopharmacy and Pharmacokinetic Properties
1998, Chemicke Listy

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An improved method for analysis of technetium radiolabeled antibody by size exclusion HPLC

Abstract

Nucl. Med. Biol.

Int. J. Nucl. Med. Biol.

Nucl. Med. Biol.

Nucl. Med. Biol.

Nucl. Med. Biol.

J. Chromatogr.

Nucl. Med. Biol.

Nucl. Med. Biol.

J. Immunol. Meth.

Optimum conditions for labeling of DTPA-coupled antibodies with 99mTc

J. Nucl. Med.

Radioimmunoscintigraphy of head and neck cancer using 99mTc-labeled monoclonal antibody E48 F(ab′)2

Cancer Res.

99mTc-human serum albumin

J. Nucl. Med.

Preclinical evaluation of an instant 99mTc-labeling kit for antibody imaging

Cancer Res.

99mTc-protein bond resistant to transchelation

Antibody Immunocoj. Radiopharm.

Directly and indirectly Tc-99m-labeled antibodies—a comparison of in vitro and animal in vivo properties

J. Nucl. Med.

Optimum conditions for labeling of DTPA-coupled antibodies with ^99mTc

Radioimmunoscintigraphy of head and neck cancer using 99mTc-labeled monoclonal antibody E48 F(ab′)₂

^99mTc-human serum albumin

Preclinical evaluation of an instant ^99mTc-labeling kit for antibody imaging

^99mTc-protein bond resistant to transchelation