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Physicochemical and Biological Characterization of RTXM83, a New Rituximab Biosimilar

  • María L. Cerutti
  • Analía Pesce
  • Cédric Bès
  • Mauricio SeigelchiferEmail author
Original Research Article

Abstract

Background

RTXM83 is a rituximab biosimilar with proven clinical safety and efficacy. It is the first rituximab biosimilar developed and approved in South America and is currently marketed in several Latin American, Middle Eastern and African countries.

Objective

The aim of this study was to present the physicochemical and biological characterization studies utilized to demonstrate the similarity between RTXM83 and its reference product.

Methods

Primary and higher order protein structures were analysed using peptide mapping with liquid chromatography–electrospray ionization–tandem mass spectrometry (LC–ESI–MS/MS), fluorescence spectroscopy and circular dichroism, and micro-differential scanning calorimetry, among other techniques. Charge variants were determined by cation-exchange chromatography (CEX) and capillary isoelectric focusing (cIEF). Glycosylation and glycoforms distribution were analysed using MS, normal phase high-performance liquid chromatography (NP-HPLC) and high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD). Size variants were evaluated by size-exclusion chromatography (SEC), sedimentation velocity analytical ultracentrifugation (SV-AUC), dynamic light scattering (DLS), and capillary electrophoresis-sodium dodecyl sulfate (CE-SDS). Biological characterization included binding assays for complement C1q, CD20, and several Fc receptors (FcRs), as well as potency determination for in vitro apoptosis induction, complement-dependent cytotoxicity (CDC), and antibody-dependent cell-mediated cytotoxicity (ADCC).

Results

RTXM83 and the reference product showed identical primary sequences and disulfide bridge patterns, and similarity at higher order protein structures, post-translational modification profiles (amino acid modifications, charge variants, and glycosylation) and levels of purity and process-related impurities. Functional studies demonstrated that RTXM83 is similar to the reference product regarding the three known mechanisms of action of rituximab: CDC, ADCC, and apoptosis induction. Binding affinities to CD20, complement component C1q, and different FcRs were also equivalent.

Conclusion

RTXM83 is similar to its reference product in all critical quality attributes.

Notes

Compliance with Ethical Standards

Funding

This study was sponsored by Mabxience.

Conflict of interest

María Laura Cerutti, Analía Pesce, Cédric Bès and Mauricio Seigelchifer are current or previous employees of Mabxience. They declare that they have no other conflicts of interest.

Supplementary material

40259_2019_349_MOESM1_ESM.pptx (5.1 mb)
Supplementary material 1 (PPTX 5241 kb)

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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • María L. Cerutti
    • 1
    • 3
  • Analía Pesce
    • 1
  • Cédric Bès
    • 2
  • Mauricio Seigelchifer
    • 1
    Email author
  1. 1.mAbxienceBuenos AiresArgentina
  2. 2.mAbxienceMadridSpain
  3. 3.Fundación Instituto Leloir-IIBBA-CONICETBuenos AiresArgentina

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