mAb Sequencing and Recombinant Expression Services

Monoclonal antibody (mAb) sequencing and recombinant expression aim to accurately decipher the amino acid sequence of monoclonal antibodies and achieve recombinant expression through genetic engineering to ensure stable supply and functional consistency. The core principle involves high-resolution sequencing of the light and heavy chains of antibodies using liquid chromatography-tandem mass spectrometry (LC-MS/MS) or next-generation sequencing (NGS). Subsequently, gene synthesis and mammalian cell expression systems (such as CHO or HEK293) are employed for recombinant antibody production, ensuring structural integrity, affinity, and biological activity.

 

mAb sequencing and recombinant expression services are widely applied in biopharmaceuticals, antibody drug development, scientific research, diagnostic reagent production, and patent protection. For instance, in antibody drug development, these services facilitate sequencing of known or unknown antibodies, followed by recombinant expression to optimize their structure and function, enhancing efficacy and stability. In the development of biosimilar antibodies, they enable sequence consistency comparison between originator and biosimilar antibodies. In research, this technology supports antibody structure-function studies and advances the development of novel antibody-based therapeutic strategies.

 



Moloney, N M. et al. Analytical Biochemistry, 2021.

Figure 1. TAFC-Specific Monoclonal Antibody Sequencing and Recombinant Expression Workflow.

 

Services at MtoZ Biolabs

Utilizing high-resolution mass spectrometry (LC-MS/MS) and next-generation sequencing (NGS) technologies, MtoZ Biolabs provides mAb sequencing and recombinant expression services to accurately determine the light and heavy chain sequences of monoclonal antibodies and achieve efficient recombinant expression through genetic engineering. Our services encompass antibody sample extraction, protein digestion, mass spectrometry sequencing, nucleic acid extraction, gene sequencing, sequence alignment, gene cloning, and stable cell line construction, ensuring high-affinity, high-stability, and biologically active recombinant antibodies.

 

Service Advantages

1. High-Precision Antibody Sequencing

MtoZ Biolabs utilizes high-resolution mass spectrometry (LC-MS/MS) and next-generation sequencing (NGS) to achieve full-sequence analysis of monoclonal antibodies, accurately identifying light and heavy chain sequences as well as post-translational modifications (PTMs), ensuring data accuracy and completeness.

 

2. Rapid and Efficient Recombinant Expression

By leveraging CHO and HEK293 mammalian cell expression systems, we optimize antibody gene cloning and stable cell line construction to achieve high-affinity, high-stability recombinant antibody expression, ensuring that the biological function remains consistent with the original antibody.

 

3. Comprehensive Bioinformatics Analysis

Utilizing advanced sequence alignment algorithms, we precisely analyze antibody gene sequences, detect mutation sites, post-translational modifications (PTMs), and optimize antibody structures, providing critical data support for antibody engineering modifications and biosimilar development.

 

4. Customized Service Solutions

MtoZ Biolabs offers flexible experimental designs tailored to different types of monoclonal antibodies (IgG, IgA, IgM, nanobodies, etc.) and various applications, including antibody drug development, patent applications, and antibody humanization, supporting antibody research and clinical translation.

 

Applications

1. Antibody Drug Development and Biosimilar Production

In the development of monoclonal antibody (mAb) drugs, mAb sequencing and recombinant expression services enable precise antibody sequence determination, structural optimization, and stable expression, ensuring biological activity and quality consistency. Additionally, these services support the development of biosimilars by optimizing manufacturing processes and improving production efficiency.

 

2. Antibody Engineering and Humanization

By sequencing the complete antibody sequence, followed by affinity maturation, Fc region modification, or humanization optimization, this service enhances antibody specificity, stability, and reduces immunogenicity, advancing targeted therapy research for cancer immunotherapy, inflammatory diseases, and autoimmune disorders.

 

3. Diagnostic Antibody Development and In Vitro Testing

In the field of in vitro diagnostics (IVD), precise monoclonal antibody sequencing and recombinant expression enable the production of high-quality antibodies with high affinity and specificity. These antibodies are essential for applications such as virus detection, cancer biomarker screening, and autoimmune disease diagnostics.

 

4. Scientific Research and Functional Studies

For fundamental research, mAb sequencing and recombinant expression services support the analysis of immune responses, antibody-antigen interactions, and structural-functional relationships. These capabilities facilitate advancements in protein engineering, vaccine development, and disease mechanism studies, driving deeper exploration in antibody science.

 

Case Study

1.Generation and Diversification of Recombinant Monoclonal Antibodies

This study aims to optimize monoclonal antibody (mAb) sequencing and recombinant expression methods to enhance efficiency in antibody engineering and biopharmaceutical development. The study subjects include monoclonal antibodies derived from hybridoma cells, B cell clones, and phage display technology. The research methodology employs high-throughput sequencing (NGS) and mass spectrometry (LC-MS/MS) to determine the sequences of antibody heavy chains (VH) and light chains (VL), followed by bioinformatics analysis to refine antibody sequences. Subsequently, gene cloning techniques are used to insert antibody genes into mammalian expression vectors, and transient or stable expression is performed in CHO or HEK293 cells to assess antibody yield, stability, and functional properties. The results indicate that the optimized sequencing method enables precise identification of variable regions, preventing expression failures due to genetic mutations or deletions. The improved expression system enhances antibody yield and stability while maintaining affinity and biological activity. Additionally, the study explores strategies for antibody sequence diversification to enhance affinity and specificity. The study concludes that integrating high-resolution sequencing with an efficient expression system enables precise antibody determination and functional optimization, providing essential technical support for antibody drug development, diagnostic reagent production, and immunotherapy.

 



DeLuca, K F. et al. eLife, 2022.

Figure 2. Recombinant Expression of Hec1 Monoclonal Antibody.

 

Deliverables

1. Comprehensive Experimental Details

 

2. Materials, Instruments, and Methods

 

3. Data Analysis, Preprocessing, and Estimation 

 

4. Bioinformatics Analysis

 

5. Raw Data Files

 

MtoZ Biolabs, an integrated Chromatography and Mass Spectrometry (MS) Services Provider, provides advanced proteomics, metabolomics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our ultimate aim is to provide more rapid, high-throughput, and cost-effective analysis, with exceptional data quality and minimal sample consumption. Free project evaluation, welcome to learn more details!