Evidence map›Paper›PMID 34643845›Full record

ArticleThe protein journal2021

Purification and Conformational Characterization of a Novel Interleukin-2 Mutein.

Leina Moro-Pérez, Sum Lai Lozada-Chang, Gabriela Rivas-García, Carlos Álvarez, Laritza Rojas-Pérez, Tammy Boggiano-Ayo, Yamile González-González

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Article in The protein journal, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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3 · Its place in the literature

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4 · The record

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5 · Who and what money

Authors and funding

7 authors.

Leina Moro-PérezBioprocess R&D Department, Center of Molecular Immunology, 216 Street and 15 Avenue, Atabey, Playa, P.O. Box 16040, 11600, Havana, Cuba. leina@cim.sld.cu.ORCID 0000-0002-2669-8131
Sum Lai Lozada-ChangBioprocess R&D Department, Center of Molecular Immunology, 216 Street and 15 Avenue, Atabey, Playa, P.O. Box 16040, 11600, Havana, Cuba.
Gabriela Rivas-GarcíaCenter of Molecular Immunology, Quality Control Direction, 216 Street and 15 Avenue, Atabey, Playa, P.O. Box 16040, 11600, Havana, Cuba.
Carlos ÁlvarezBiology Faculty, Center for Protein Studies, University of Havana, 25 St. No. 455, Vedado, 10400, Havana, Cuba.
Laritza Rojas-PérezBiology Faculty, Center for Protein Studies, University of Havana, 25 St. No. 455, Vedado, 10400, Havana, Cuba.
Tammy Boggiano-AyoBioprocess R&D Department, Center of Molecular Immunology, 216 Street and 15 Avenue, Atabey, Playa, P.O. Box 16040, 11600, Havana, Cuba.
Yamile González-GonzálezCenter of Molecular Immunology, EPOVAC Facility, 216 Street and 15 Avenue, Atabey, Playa, P.O. Box 16040, 11600, Havana, Cuba.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Toxicity of high-dose IL-2-based therapies have motivated the development of the IL-2 mutein, which has low expansion properties for regulatory T lymphocytes. The development of two variants (A and B) for the IL-2 mutein purification as well as a conformational comparative study by Circular dichroism (CD) and fluorescence spectroscopy of these products were evaluated. For the first time, in our center, were used of DTT and 2% SDS in the solubilization step to decrease the aggregates on intermediate product, which favors that disulfide bridges are correctly formed during re-folding. A molecular weight of 18 kDa to the monomeric form and of 25-37 kDa to the oligomeric species were estimated by SDS-PAGE. IL-2 mutein showed similar far-UV CD spectral characteristic typical of cytokines with 41% of α-helix content. Batches obtained by Process B showed similar conformational features according near-UV CD and FS studies. However, those obtained by Process A differed in their folding. IL-2 mutein showed that conformational features by near-UV CD were affected by 2% SDS, no variations on secondary structure were observed. Melting temperature values by far-UV CD were higher than 95 °C, indicating a high thermal stability. Finally, the drug product obtained by Process B showed similar conformational characteristics by near-UV CD and FS, and higher biological activity values (7.0 × 10

Indexed as

Interleukin-2Circular DichroismElectrophoresis, Polyacrylamide GelProtein Structure, SecondaryTemperatureInterleukin-2Cell proliferation assayCharacterizationCircular dichroismFluorescence spectroscopyIL-2 muteinPurification

Identifiers

PMID34643845

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