Technical Service

Mammalian Expression System

Mammalian Expression System

  The mammalian cell protein expression system can perform post-translational modification and folding during the expression of proteins. Thus, the protein expressed by mammalian cell expression system has a similar spatial structure and modification to the wild type protein, which resulting in biological activity. Mammalian cell protein expression system is widely applicable in producing functional proteins, antibodies and clinic vaccines.

AtaGenix Advantages

1. AtaGenix's success rate is up to 90%.  

2. AtaGenix provides a variety of expression systems: transient transfection, recombinant antibody expression and production, stable cell line expression service, etc.  

a) Transient transfection expression: Multiple cell lines are cultured in serum free medium (three 293 suspension cells and three CHO suspension cells) are offered, and our highly efficient expression vectors, which we own the independent intelligent property rights, can raise the yield to 3~6 fold. In terms of recombinant antibodies, we developed a highly efficient vector-cell-transfection system by repeated identification  

b) Stable cell line construction: AtaGenix owns the independent intelligent property rights of a modified CHO-K1 cell line, which is used for protein and antibody research and production for clients  

3. Large-scale production: the serum-free culture cells can reach up to 200L per batch.  

4. One-stop service: Design and optimize gene, transient transfection, stable cell line construction, recombinant antibody production and management of data and report.

service content

Transient expression service

Service Name

Service Content

Leading Time

Selection of expression system

Expression vector: Commercial vector, modified highly efficient vector that was improved by AtaGenix.

Expression cell line: Multiple cell lines cultured in serum free medium (three 293 suspension cells and three CHO suspension cells)

 

Construction of the plasmid

Gene optimization, gene synthesis & subclone

2-4weeks

Expression & Purification test

Endotoxin-free plasmid extraction, plasmid quality control and transient transfection, optimize the expression condition, SDS-PAGE/WB assay, 80ml amplification and purification test on the optimum conditions, tag removing test (optional).

2-3weeks

1-10L Expression & Purification

Endotoxin-free plasmid extraction in a large amount, plasmid quality control and transient transfection, Protein affinity purification (or other purification method), protein buffer replacement and concentration, SDS-PAGE/WB identification of the purified protein, MS identification (optional), tag removing (optional)

2-4weeks

Large-scale fermentation & purification

Endotoxin-free plasmid extraction in a large amount, plasmid quality control and transient transfection, Protein affinity purification (or other purification method), protein buffer replacement and concentration, SDS-PAGE/WB identification of the purified protein, MS identification (optional)

10-20L

3-5 weeks

20-50L

4-7 weeks

Above 50L

5-12 weeks



Stable cell line construction service

Service name

Service contents

Lead time

Selection of expression system

Expression vector: Commercial vector, Expression cell line: Commercialized cell lines and the modified CHO-K1 cell line which AtaGenix owns independent intelligent property right

 

Construction of the plasmid

Gene optimization and design, gene synthesis & subclone

2-4 weeks

Stable cell line construction

Endotoxin-free plasmid extraction, plasmid quality control and transient transfection (293 & CHO), expression test, SDS-PAGE/WB assay, screening by pressure, cell subclone, expression and adaption evaluate under serum free circumstance, build library of cell line.  

2-4 months

1-10L Expression & Purification

Protein buffer replacement and concentration, SDS-PAGE/WB identification of the purified protein, MS identification (optional), tag removing (optional)

2-4 weeks

Large-scale fermentation & purification

Protein affinity purification (or other purification method), protein buffer replacement and concentration, SDS-PAGE/WB identification of the purified protein, MS identification (optional).

10-20L

3-5 weeks

20-50L

4-7 weeks

Above 50L

5-12 weeks

Case studyA

  In this case, AtaGenix was requested to produce Protein H using our custom 1L protein expression. Protein H is expressed by the secretory system. We tested pATX1 and commercialized pcDNA3.1, and the results showed that protein H does not express in pcDNA3.1 vector, however it was expressed successfully in pATX1. The amount was 4.53mg/L after purification. We delivered the protein we had obtained as our client required.


哺乳动物细胞表达案例1

哺乳动物细胞表达案例2

哺乳动物细胞表达案例3

Case B

  Protein L expressed successfully in pATX2, the molecular weight is 47.1kDa theoretically, and it has two glycosylation site. Thus, the actual molecular weight is 55kDa post expression and the amount produced was 247.5mg/L after purification

哺乳动物细胞表达案例4

哺乳动物细胞表达案例5

哺乳动物细胞表达案例6


Consultation and ordering:

Tel: 0086-27-8743 3958

Email:info@atagenix.com