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Evaluation of Iminodiacetic Acid (IDA) as an Ionogenic Group for Adsorption of IgG1 Monoclonal Antibodies by Membrane Chromatography

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Abstract

Iminodiacetic acid (IDA) is one of the chelating ligands most frequently employed in immobilized metal-ion affinity chromatography (IMAC) due to its ability to act as electron-pair donor, forming stable complexes with intermediate and borderline Lewis metal ions (electron acceptor). Thus, IDA can also be employed in ion exchange chromatography to purify positively charged proteins at neutral pH values. This study aimed to evaluate IDA as an ionogenic group (ion exchanger) immobilized on poly (ethylene vinyl alcohol) (PEVA) hollow fiber membranes for immunoglobulin G1 (IgG1) monoclonal antibody (MAb) purification. IDA-PEVA membranes showed considerable promise for MAb purification, since IgG1 was recovered in eluted fractions with traces of contaminants as confirmed by Western blotting and ELISA analysis. Quantification of IgG1 showed that a purity of 94.2% was reached in the elution step. Breakthrough curve and batch adsorption experiments showed that the MAb dynamic binding capacity (DBC) of 3.10 mg g−1 and the maximum adsorption capacity of 70 mg g−1 were of the same order of magnitude as those found in the literature. The results obtained showed that the IDA-PEVA hollow fiber membrane could be a powerful adsorbent for integrating large-scale processes for purification of MAb from cell culture supernatant.

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Acknowledgments

The authors are grateful to Dr. W.M.S.C. Tamashiro and Dr. E.F.P. Augusto for providing the cell supernatant and assistance with the ELISA tests.

Funding

This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo, FAPESP, Brazil (grant number 2004/09896-8) and a MSc fellowship from Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq, Brazil (grant number 141930/2006-3). It was also partially financed by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brazil (CAPES), Finance Code 001.

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Authors and Affiliations

  1. Chemical Engineering Department, Federal University of São Paulo (UNIFESP), Diadema, SP, 09913-030, Brazil

    Igor Tadeu Lazzarotto Bresolin & Iara Rocha Antunes Pereira Bresolin

  2. School of Chemical Engineering, University of Campinas (UNICAMP), Campinas, SP, 13083-970, Brazil

    Sônia Maria Alves Bueno

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  1. Igor Tadeu Lazzarotto Bresolin
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Correspondence to Igor Tadeu Lazzarotto Bresolin.

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Bresolin, I.T.L., Bresolin, I.R.A.P. & Bueno, S.M.A. Evaluation of Iminodiacetic Acid (IDA) as an Ionogenic Group for Adsorption of IgG1 Monoclonal Antibodies by Membrane Chromatography. Appl Biochem Biotechnol 191, 810–823 (2020). https://doi.org/10.1007/s12010-019-03217-5

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