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Antibody Sequencing Services

Tek Biotech provides rapid and professional antibody sequencing services (including hybridoma sequencing, full-length antibody sequencing, etc.) to sequence the genes encoding the heavy chain variable (VH) region and light chain variable (VL) region of monoclonal antibodies against hybridoma cells. Meanwhile, Tek Biotech is equipped with advanced antibody full-length sequencing technologies, such as liquid chromatography (LC-MS/MS) coupled with tandem mass spectrometry (MS/MS) and supporting advanced computational algorithms, to provide accurate and rapid antibody full-length sequencing services according to your needs.

█ Hybridoma Sequencing Service

Tek Biotech can complete high-throughput (NGS) hybridoma sequencing in 4-6 weeks and provide the customer with all VH and VL sequences as well as a full report informing the customer of the sequence of the monoclonal antibody to the hybridoma cells (including sequence analysis of the CDR region of the monoclonal antibody relative to the VH and VL sequences) and any heterogeneity or other antibody isoforms that may be present.

After hybridoma sequencing, the corresponding sequence can be obtained, and when monoclonal antibodies are needed in subsequent projects, the measured gene sequence can be used for the preparation of recombinant antibodies. It solves the problem of cell death or loss of high specificity in the process of hybridoma preservation. Hybridoma sequencing is also important for further large-scale production and humanization of antibodies and other bioengineering operations.

█ Hybridoma Sequencing Service Process

Project

Content

Deliveries

Period

Hybridoma Cell Antibody Sequencing Service

RNA Extraction

Reverse transcription

PCR amplification of VH and VL sequences

Sequencing validation

Gene synthesis

Expression vector construction

Antibody VH and VL sequences

Expression vectors containing VH and VL genes, respectively (plasmid construct QC)

2-3 weeks

█ Service Advantages

-- Applicable to any species

-- Uses random primers that do not introduce mutations

-- Identifies all VH and VL sequences and quantifies their relative abundance

-- Absolute sequence confidentiality

-- High-throughput approach allows simultaneous sequencing of up to 50 hybridomas -- Bulk hybridoma sequencing package available

-- Sequences delivered within 3-4 weeks

█ Antibody Full-length Sequencing

There are many monoclonal antibodies for which hybridoma cell lines are not available. In order to satisfy customers' sequencing requirements for unknown antibody sequences (no hybridoma cell lines), Tek Biotech Biologics also offers full-length sequencing of antibodies, which determines the amino acid sequence of variable domains by combining mass spectrometry with ab initio peptide sequencing and overlapping peptide assembly. Full-length sequencing of antibodies is the analytical process of deducing the amino acid sequence of an antibody and discovering any associated post-translational modifications without prior knowledge of the DNA or protein sequence.

Tek Biotech guarantees functional antibody sequences and comprehensive sequencing reports with complete sequence coverage. Based on the Recombinant Antibody Transient Expression Service, Tek Biotech can sequence an antibody and deliver the corresponding recombinant protein in as little as 9 weeks.

█ Sample Request

Sample size: 100μg monoclonal antibody protein

Concentration: >0.2mg/mL

Purity: 95% or more of the total protein amount of the target antibody

Others: no antibody protein of the same antigen

█ Service Content

Mass Spectrometry Sequencing

Antibody Expression

Period

Eeliveries

Digested into overlapping peptides;

LC-MS/MS using Thermo Orbitrap

Ab initio identification of peptides;

Peptide assembly;

Gene synthesis

Cloning

Transient expression in HEK293 cells

Affinity Purification

Buffer exchange (PBS)

Sterile filtration

Purity >98% (SDS-PAGE)

Endotoxin < 1 EU/mg

3-4 weeks

Comprehensive Sequencing report;

>2 mg purified antibody;

QC report

█ Service Advantages

-- Fast: Sequencing can be completed within 10 business days

-- High-throughput: our DASS methods are high-throughput compatible

-- Specialized: scientists are responsible for each project with a dedicated team of professionals for quality control at every step of the process; expert R&D teams work to optimize protocols to deliver higher quality data

-- Personalization: Customized services include, but are not limited to, antibody design, cloning, ab initio sequencing and analysis, and in vitro optimization to meet your needs.

-- High accuracy: strong peak signatures are obtained in multiple mass spectrometry reads for individual peptides, correctly identifying each amino acid in the sequence.

-- Wide range of applications: can be used for all types of antibodies (except human). It has even been used to sequence contaminated or bound antibodies or pairs of antibodies with different light chains.

-- Informative: Ab initio sequencing of antibodies often results in the detection of unexpected differences in sequencing or new glycosylation sites on protein molecules that cannot be detected by DNA sequencing.

Antibody Sequencing Services Frequently Asked Questions

Q: During the sequencing process, contamination or hybridization may occur, resulting in inaccurate sequence results.

A: Strictly control experimental operations, adopt aseptic techniques and strict laboratory operating procedures; Regularly inspect the cleanliness and disinfection of instruments and equipment; Avoid cross contamination and hybridization during sample processing and operation.
Q: The obtained sequence has low quality, including insufficient sequence length, base errors, or low sequencing quality.

A: Optimize experimental conditions and sequencing methods, including optimizing PCR reaction conditions, using high-quality primers, selecting appropriate sequencing platforms, etc; Strengthen the quality control of DNA samples, such as DNA purification, modification, and concentration processing steps.
Q: Deviation occurs during PCR amplification, resulting in over amplification or non amplification of some sequences.

A: Optimize PCR reaction conditions, including adjusting parameters such as temperature, primer concentration, and number of cycles; Using high-quality DNA templates and appropriate PCR enzymes; Optimize the amplification conditions and loop program to avoid causing amplification bias.
Q: There are variations or mutations in the gene sequence that affect the accuracy and reliability of sequencing results.

A: Perform sequence alignment and analysis, identify and distinguish true variations and mutations, as well as sequencing errors; Conduct validation experiments, such as repeated sequencing, using different sequencing platforms or methods, to confirm the accuracy of mutations and mutations; Revise and correct the sequence results to ensure their accuracy and reliability.

Q: During the sequencing process, contamination or hybridization may occur, resulting in inaccurate sequence results.

A: Strictly control experimental operations, adopt aseptic techniques and strict laboratory operating procedures; Regularly inspect the cleanliness and disinfection of instruments and equipment; Avoid cross contamination and hybridization during sample processing and operation.
Q: The obtained sequence has low quality, including insufficient sequence length, base errors, or low sequencing quality.

A: Optimize experimental conditions and sequencing methods, including optimizing PCR reaction conditions, using high-quality primers, selecting appropriate sequencing platforms, etc; Strengthen the quality control of DNA samples, such as DNA purification, modification, and concentration processing steps.
Q: Deviation occurs during PCR amplification, resulting in over amplification or non amplification of some sequences.

A: Optimize PCR reaction conditions, including adjusting parameters such as temperature, primer concentration, and number of cycles; Using high-quality DNA templates and appropriate PCR enzymes; Optimize the amplification conditions and loop program to avoid causing amplification bias.
Q: There are variations or mutations in the gene sequence that affect the accuracy and reliability of sequencing results.

A: Perform sequence alignment and analysis, identify and distinguish true variations and mutations, as well as sequencing errors; Conduct validation experiments, such as repeated sequencing, using different sequencing platforms or methods, to confirm the accuracy of mutations and mutations; Revise and correct the sequence results to ensure their accuracy and reliability.

Consult Now Antibody Sequencing Services

Antibody Detection Platform

█ Hybridoma Sequencing Service

█ Hybridoma Sequencing Service Process

█ Service Advantages

█ Service Advantages

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Antibody Sequencing Services Frequently Asked Questions

Q: During the sequencing process, contamination or hybridization may occur, resulting in inaccurate sequence results.

Q: The obtained sequence has low quality, including insufficient sequence length, base errors, or low sequencing quality.

Q: Deviation occurs during PCR amplification, resulting in over amplification or non amplification of some sequences.

Q: There are variations or mutations in the gene sequence that affect the accuracy and reliability of sequencing results.

Q: During the sequencing process, contamination or hybridization may occur, resulting in inaccurate sequence results.

Q: The obtained sequence has low quality, including insufficient sequence length, base errors, or low sequencing quality.

Q: Deviation occurs during PCR amplification, resulting in over amplification or non amplification of some sequences.

Q: There are variations or mutations in the gene sequence that affect the accuracy and reliability of sequencing results.

Consult Now Antibody Sequencing Services