Antibody Light and Heavy Chain Sequencing Service

Antibodies consist of light chains (LC) and heavy chains (HC), and these two parts together determine the specificity and function of the antibody. Through efficient sequencing technology, we can precisely resolve the full-length amino acid sequences of LC and HC, providing solid data support for antibody development and application.

 

MtoZ Biolabs has developed an efficient, fast, and accurate antibody light and heavy chain sequencing platform based on high-throughput mass spectrometry (MS) and PCR technology. Our team of experienced scientists and technical experts has deep expertise in antibody light and heavy chain and its sequencing research. We are committed to providing high-quality, efficient services to advance antibody light and heavy chain sequencing research and antibody-related drug development. Free project evaluation, welcome to learn more details! Our technical specialists are available to provide a free business assessment.

 

Technical Principles

Antibody sequencing technology mainly includes PCR and MS analysis.

 

1. PCR Amplification

(1) Primer Design: Designing specific primers based on known sequences or conserved regions of LC and HC.

(2) RNA Extraction and Reverse Transcription: Extracting total RNA from antibody-producing cells and reverse transcribe RNA into cDNA using reverse transcriptase.

(3) PCR Amplification: Using designed primers to perform PCR amplification on cDNA to obtain specific amplification products of LC and HC.

 

2. Mass Spectrometry Analysis

(1) Sample Purification: Purifying PCR products to remove impurities.

(2) Enzymatic Digestion: Using specific proteases (such as trypsin) to cleave purified antibodies into smaller peptides.

(3) MS Analysis: Performing ionization and mass analysis of the peptides using High-Resolution Mass Spectrometry (HRMS) and Tandem Mass Spectrometry (MS/MS) technology.

(4) Data Interpretation: Using specialized bioinformatics tools to interpret MS data and reconstruct the amino acid sequences of LC and HC.

 

Analysis Workflow

1. Sample Preparation

 (1) Extracting total RNA from antibody-producing cells and reverse transcribe into cDNA.

 (2) Designing and synthesize specific primers.

 

2. PCR Amplification

 Performing PCR amplification of specific fragments of LC and HC.

 

3. Mass Spectrometry Analysis

 (1) Purifying PCR amplification products.

 (2) Using trypsin to enzymatically digest the purified products into peptides.

 (3) Performing ionization and mass analysis of the peptides using HRMS and MS/MS technology.

 

4. Data Interpretation

Using specialized software to reconstruct the full-length amino acid sequences of LC and HC.

 

5. Validation and Reporting

(1) Validating sequencing results through further experiments.

(2) Generating a detailed sequencing report, including the complete amino acid sequences of LC and HC.

 

Service Advantages

1. High Accuracy

We can ensure accurate resolution of amino acid sequences using HRMS and MS/MS technology.

 

2. High Throughput

Our platform is able to sequence multiple samples simultaneously, improving research efficiency.

 

3. Comprehensive Coverage

Our service covers complete resolution of the full-length sequences of LC and HC, providing comprehensive structural information.

 

4. Customized Service

We provide customized sequencing solutions and full technical support to meet our clients' needs.

 

Applications

1. Discovering Novel Antigen-specific Monoclonal Antibodies

  



Devulapally, PR. et al. Genome Med. 2018.

 

2. Personalized Medicine

  



Dupré, M. et al. Anal Chem. 2021.

 

Deliverables

1. Experimental Procedures

2. Relevant Mass Spectrometry Parameters

3. Detailed Information on Antibody Light and Heavy Chain Sequencing

4. Mass Spectrometry Images

5. Raw Data