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Address:Room 502, Building B15, Wuqing Developmental Area, Tianjin, China

Email:info@kmdbioscience.com

Protein Sequencing Service

Protein primary sequence determines the advanced structure of a protein, influencing a protein’s function. Therefore, the sequence of protein is important to study the biological function of the protein. KMD Bioscience has developed a comprehensive protein sequencing platform to meet various needs of our clients, including N-terminal and de novo sequencing. Our featured services are presented below, but if you have any special requirement, you are welcome to contact us for custom service.

 

De Novo Sequencing Service

Based on the most advanced mass spectrometry instrument, ‘Dao Jin‘ PPSQ-31A, and combined with rich experience in bioinformatics analysis, KMD Bioscience has established a whole new generation of de novo sequencing platform that can achieve accurate sequence analysis of monoclonal antibodies and proteins. After receiving the sample, we will first identify and fragment it with 6 commonly used proteases: Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C and Lys-C. Our goal is to get the most fragmented sequences to achieve 100% protein coverage. The sequence information was spliced by PEAKs Studio and PEAKs Ab software, and then artificially spliced to achieve nearly 100% correct protein or antibody primary sequence.

* For more de novo sequencing services, please consult the antibody de novo sequencing services or contact our technicians for further information.

 

N-Terminal Sequencing Service (Edman Degradation)

We can sequence up to 60 amino acid residues from the N-terminal. The Edman method for sequencing a single amino acid cycle consists of 3 steps.

Step 1: PITC binds to free amino acids at the N-terminal of the protein under alkaline conditions.

Step 2: N-terminal residues are removed in acidic solutions.

Step 3: PITC-bound residues are converted to more stable PTH residues. After on-line analysis by HPLC, amino acids were determined according to elution time.

 

Sequencing Procedure

Protein samples were first separated by SDS-PAGE to ensure the purity can meet the requirements of sequencing. Subsequently, protein samples from SDS-PAGE were transferred to PVDF membranes and cut the gel band after dyeing. Then loaded it directly on the sequencer.

 

Sample Requirement:

1. Purity: >95 (Based on moles).

2. Sample amount: 50-100pmol (Liquid volume control at 20ul).

3. Unblocked N-terminus.

4. Sample format: Dry, solution, PVDF membranes.

5. No interference: Protease, amino acid, amine, salt, acetaldehyde, detergent, etc.

* For specific sample requirements and examples, please click on the ‘Edman Sequencing Sample Requirements’ for more information.

 

Service Characteristics

* Identify recombinant protein expression and pulldown products.

* Validation of expressed products in cell lines.

* Validation of N-terminal sequences of antibody drugs.

 

Service Highlights

* Unparalleled protein sequencing accuracy.

* 100% protein sequence coverage and confidence.

* Strict quality control system and strong analytical capabilities.