Comparison of two-dimensional gel electrophoresis patterns and MALDI-TOF MS analysis of therapeutic recombinant monoclonal antibodies trastuzumab and rituximab

Nebija, Dashnor; Kopelent‐Frank, H.; Urban, Ernst; Noe, Christian R.; Lachmann, Bodo

doi:10.1016/j.jpba.2011.07.006

Cited by 21 publications

(23 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…2D-gel electrophoresis is particularly well suited for this purpose, due to the option for direct visual pattern recognition and the option for easy additional analysis. Recently, we investigated this method for the analysis of monoclonal antibodies trastuzumab and rituximab [2]. In the present paper, we report our follow-up work on trastuzumab, particular in view of the suitability of the method for stability studies.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Quality Control and Stability Studies with the Monoclonal Antibody, Trastuzumab: Application of 1D- vs. 2D-Gel Electrophoresis

Nebija¹,

Noe

Urban

et al. 2014

IJMS

Self Cite

View full text Add to dashboard Cite

Recombinant monoclonal antibodies (rmAbs) are medicinal products obtained by rDNA technology. Consequently, like other biopharmaceuticals, they require the extensive and rigorous characterization of the quality attributes, such as identity, structural integrity, purity and stability. The aim of this work was to study the suitability of gel electrophoresis for the assessment of charge heterogeneity, post-translational modifications and the stability of the therapeutic, recombinant monoclonal antibody, trastuzumab. One-dimensional, SDS-PAGE, under reducing and non-reducing conditions, and two-dimensional gel electrophoresis were used for the determination of molecular mass (Mr), the isoelectric point (pI), charge-related isoform patterns and the stability of trastuzumab, subjected to stressed degradation and long-term conditions. For the assessment of the influence of glycosylation in the charge heterogeneity pattern of trastuzumab, an enzymatic deglycosylation study has been performed using N-glycosidase F and sialidase, whereas carboxypeptidase B was used for the lysine truncation study. Experimental data documented that 1D and 2D gel electrophoresis represent fast and easy methods to evaluate the quality of biological medicinal products. Important stability parameters, such as the protein aggregation, can be assessed, as well.

show abstract

Section: Introductionmentioning

confidence: 99%

“…Trastuzumab is composed of 1328 amino acids. It contains two identical heavy (HC) and two identical light chains (LC) linked via disulfide bonds [2,5]. Each HC contains 450 amino acids, and each LC contains 214 amino acids.…”

Section: Introductionmentioning

confidence: 99%

Quality Control and Stability Studies with the Monoclonal Antibody, Trastuzumab: Application of 1D- vs. 2D-Gel Electrophoresis

Nebija¹,

Noe

Urban

et al. 2014

IJMS

Self Cite

View full text Add to dashboard Cite

show abstract

“…4 Amongst several potential PTMs of amino acids, deamidation of asparagine or glutamine and oxidation of methionine or tryptophan represent common chemical modifications for mAbs during downstream processing and storage. Figures 6A and 6B show the extent of amino acid oxidation, and deamidation, respectively, for oxidations for the different digestion methods.…”

Section: Resultsmentioning

confidence: 99%

A Method for Mapping Glycosylation Sites in Proteins

Han

Simpson

2017

J Biomol Tech

View full text Add to dashboard Cite

“…These typical studies include antibody primary structure by chemical sequencing (Edman degradation) as well as peptide mapping by means of ESI-MS and MALDI-MS [26][27][28]; higher order structure by RP-HPLC-ESI-MS, Ellman's assay CD, FTIR, hydrogen deuterium exchange (HDX)-MS and X-ray [28,29]; post-translational modifications (PTM) and charge variants by MSion-exchange chromatography (IEC), hydrophobic interaction chromatography (HIC), capillary electrophoresis (CE) and isoelectric focusing (IEF) [30]; glycan profile and variants and size heterogeneity by HPLC-fluorescence, size-exclusion chromatography (SEC), native gel, capillary electrophoresis, RP-HPLC-MS and native intact MS [31], as well as solubility measurement by ultrafiltration or PEG-induced precipitation methods [32,33]. Moreover, the immunogenicity should be evaluated for both antibody and ADCs [34,35], but unnecessary for SMs.…”

Section: General Differences Between Sms and Lms (Mab And Adc) In Admmentioning

confidence: 99%

An Overall Comparison of Small Molecules and Large Biologics in ADME Testing

Wan

2016

ADMET DMPK

View full text Add to dashboard Cite

Biologics mainly monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) as new therapeutics are becoming increasingly important biotherapeutics. This review is intended to provide an overall comparison between small molecules (SMs) and biologics or large molecules (LMs) concerning drug metabolism and pharmacokinetic (DMPK) or associated with absorption, distribution, metabolism and elimination (ADME) testing from pharmaceutical industry drug discovery and development points of view, which will help design and conduct relevant ADME testing for biologics such as mAbs and ADCs. Recent advancements in the ADME for testing biologics and related bioanalytical methods are discussed with an emphasis on ADC drug development as an example to understand its complexity and challenges from extensive in vitro characterization to in vivo animal PK studies. General non-clinical safety evaluations of biologics in particular for ADC drugs are outlined including drug-drug interaction (DDI)and metabolite/catabolite assessments. Regulatory guidance on the ADME testing and safety evaluations including immunogenicity as well as bioanalytical considerations are addressed for LMs. In addition, the preclinical and human PK data of two marked ADC drugs (ADCETRIS, as examples are briefly discussed with regard to PK considerations and PK/PD perspectives.

show abstract

Comparison of two-dimensional gel electrophoresis patterns and MALDI-TOF MS analysis of therapeutic recombinant monoclonal antibodies trastuzumab and rituximab

Cited by 21 publications

References 36 publications

Quality Control and Stability Studies with the Monoclonal Antibody, Trastuzumab: Application of 1D- vs. 2D-Gel Electrophoresis

Quality Control and Stability Studies with the Monoclonal Antibody, Trastuzumab: Application of 1D- vs. 2D-Gel Electrophoresis

A Method for Mapping Glycosylation Sites in Proteins

An Overall Comparison of Small Molecules and Large Biologics in ADME Testing

Contact Info

Product

Resources

About