Antibody Characterization Service

Antibody characterization encompasses comprehensive analysis of structural, physicochemical, immunological, and biological properties of antibody, along with impurity assessment and precise antibody quantification. It not only provides a reliable basis for clinical trials but also serves as a critical foundation for the commercialization of antibodies as therapeutic drugs. Through characterization, a thorough understanding of antibody functionality, stability, and safety can be achieved, enabling optimization of product consistency and quality control to meet stringent regulatory requirements. Furthermore, antibody characterization supports impurity elimination, precise dosage adjustment, and efficacy prediction, accelerating drug development, enhancing market competitiveness, and strengthening patient trust.

 

Service at MtoZ Biolabs

MtoZ Biolabs is committed to becoming a leading provider of mass spectrometry-based proteomics services, leveraging years of expertise in proteomics to deliver high-quality antibody characterization service to global clients. The company is equipped with a wide range of advanced instruments, including state-of-the-art mass spectrometers, Nano-LC systems, capillary electrophoresis instruments, and dynamic light scattering (DLS) devices, covering various needs from molecular weight determination to structural characterization. Combined with cutting-edge technology, we achieve high sensitivity and high-resolution data acquisition in antibody characterization. Our antibody characterization services include structural characterization, physicochemical property analysis, immunological property assessment, biological activity testing, impurity evaluation, and antibody quantification. Through systematic and multidimensional analysis, we provide comprehensive data that comply with international regulatory standards, accelerating drug development and commercialization. MtoZ Biolabs is dedicated to offering comprehensive and reliable technical support to antibody developers, ensuring the functionality, safety, and consistency of antibody products, and contributing to global biopharmaceutical innovation.

 

Service Advantages

1. High Specificity and Consistency Assurance: MtoZ Biolabs utilizes advanced mass spectrometry techniques and rigorous validation processes to ensure antibodies accurately recognize target molecules while effectively avoiding non-specific binding. This fundamentally enhances research reliability and reproducibility, reducing resource waste and project delays.

 

2. State-of-the-Art Equipment and Expertise: MtoZ Biolabs is equipped with cutting-edge instruments and able to address the challenge of insufficient characterization data for most antibodies, providing researchers with comprehensive data support.

 

3. Batch Consistency Verification: We conduct stringent inter-batch consistency testing to ensure the stability and reproducibility of biological reagents. This resolves the quality inconsistency issues caused by batch-to-batch variability, safeguarding the continuity and reliability of research projects.

 

4. Customized Solutions and Global Support: Tailored to the specific research needs of clients, we offer flexible antibody characterization solutions, including validation of specific target molecules and assessment of batch variability. Our data complies with international regulatory standards, providing reliable technical support and fast service responses to global biopharmaceutical customers.

 

Case Study

1. Specificity Testing of 24 mAbs by Hydrophobic Interaction Chromatography

Researchers used hydrophobic interaction chromatography (HIC) to determine the hydrophobicity of monoclonal antibodies (mAbs) for antibody identification and compared the apparent HIC retention factors (k) of 24 mAbs. Due to the size differences between fusion proteins, bispecific antibodies (bsAb), and the high heterogeneity of antibody-drug conjugates (ADC), only mAb products were injected into HIC to study the impact of protein hydrophobicity on size exclusion chromatography (SEC) elution times. The HIC retention factors ranged from 1 to 6, showing significant differences in hydrophobicity among the 23 FDA and EMA-approved mAb products. Fourteen mAbs had k values between 1 and 3, considered moderately hydrophobic, while eight mAbs had k values between 3 and 5, classified as moderately to highly hydrophobic. When the HIC retention factor exceeded 5, atezolizumab and pembrolizumab were found to be the two most hydrophobic mAbs. Non-glycosylated atezolizumab and pembrolizumab are expected to exhibit such high hydrophobicity.

 

 

 Goyon, A. et al. J Chromatogr B Analyt Technol Biomed Life Sci. 2017.

 

2. Identification of a Monoclonal Antibody Clipping Variant by Cross-Validation Using Capillary Electrophoresis – Sodium Dodecyl Sulfate, Capillary Zone Electrophoresis – Mass Spectrometry and Capillary Isoelectric Focusing – Mass Spectrometry

Based on the CZE-MS analysis of the reduced subunits, it can be concluded that the loss of 105 amino acids occurs on the heavy chain. However, an important question remains regarding whether this clipping happens at the N-terminal of the heavy chain. To address this, the enzyme IdeS was chosen for further analysis, as IdeS cleaves IgG right after its hinge region. Both non-reduced and reduced IdeS subunit analyses were conducted. The non-reduced IdeS analysis clearly shows a F(ab′)2 clipping variant with a mass of 87,116.57 Da, which is very close to the theoretical molecular weight (87,114.33 Da) of the loss of 105 amino acids in the F(ab′)2. In the reduced IdeS subunit CZE-MS analysis, the clipping variant is observed near the end of the migration (P6 in Fig. 7A). This clipping variant is identified as an Fd′ clipping variant, with a mass of 14,287.90 Da, very close to the theoretical molecular weight (14,288.13 Da) for the loss of 105 amino acids in the Fd′. Additionally, post-translational modifications (PTMs) on the reduced IdeS subunits were also observed. In P2, a mass of 25,250.42 Da is related to oxidation and deamidation on Fc/2, while deamidation on LC is detected in P4. CE-MS was used to directly confirm the molecular weight of an unknown peak in the CE-SDS. The analysis of non-reduced and DTT-reduced IdeS digests of bevacizumab provides more in-depth information. This subunit-oriented approach accurately locates the clipping variant on the antibody heavy chain N-terminal. Through this process, the antibody characterization is fully elucidated, including the identification of antibody variants.

 

Li, M. et al. J Chromatogr A. 2022.

 

Applications

1. Quality Control by Antibody Characterization

 



Evans, AR. et al. MAbs. 2024.

 

FAQ

1. How to improve specificity and sensitivity of antibody?

Rigorous validation processes and multiple technical approaches, such as western blot, immunoprecipitation, immunofluorescence, and mass spectrometry, can be employed for comprehensive antibody evaluation. These measures ensure that antibodies accurately recognize their target molecules, minimize non-specific binding and background signals, and enhance the accuracy and reliability of experimental results.

 

Deliverables

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. Relevant Liquid Chromatography and Mass Spectrometry Parameters

4. The Detailed Information of Antibody Characterization 

5. Mass Spectrometry Image

6. Raw Data