Welcome to TGpeptide-Professional peptide manufacturer.

Service hotline:+86-(0)-18115476705

Mammalian expression

     Mammalian cells can correctly and effectively express eukaryotic proteins, including the synthesis, processing, and secretion of proteins. The mammalian system has the excellent function of post-translational modification of proteins and can assemble proteins correctly. Our Mammalian expression can provide flexible services which could meet different custom needs, including tag removal, endotoxin control, SEC-HPLC assay, glycosylation analysis and so on.

  • Transient Transfection

          Mammalian cells are capable of protein folding and post-translational modification, which can express proteins with molecular structures, physical and chemical structures, and biological functions mostly approaching natural higher-orgainism proteins. It brings more possibility to obtain the same bioactivity as natural protein. Cells that are frequently used for transient transfection are HEK293 or CHO cells. Large volume transient transfection of HEK293 and CHO cells adapted to suspension culture has addressed the need to obtain amounts of recombinant protein without the time-consuming and labor-intensive creation of stable cell lines.

Service procedures

Specification

Timeline

Gene synthesis and codon optimization (optional)

         

1-2 weeks

Vector construction

clone cDNA into expression vector

plasmid sequencing

large-scale/plasmid preparation

1-2 weeks

Expression and purification

transient transfection of HEK293/CHO cells

protein purification

SDS-PAGE and UV analysis

2-3 weeks

Large-scale expression and purification (optional)

2-3 weeks

Deliverables:

Sequencing report


Feasibility report


0.1-0.5 mg sample (If expression is feasible and sample remains from QC testing)


Purified protein


SDS-PAGE report


COA


  • Stable Transfection

          Stable cell line is widely used in various areas, such as recombinant protein and antibody production, testing and analysis, gene editing, functional study and so on. Engaged in protein expression study for years and possessing abundant stable cell line development experience, TGpeptide can increase stable cell line productivity and improve experimental results.

Service procedures

Specification

Timeline


Testing

Host cell line preparation

Killing curve test

/

Plasmid construct

Codon optimization and gene synthesize

Subcloning

Max-prep target plasmid in transfection grade

3 weeks

Transfection

Electroporation

Nucleofector

1 week

Screening

Mark up the single clones/cell pools and check the titers

Screen and scale up the positive clones (single clones/cell pools )

cell pools: 4weeks
single clones: 8weeks

Amplification(optional)

MTX/MSX amplification

Subcloning

Screen single clones

2-3 months

QC

tability test stability test for 10-50 passages

Batch culture

Mycoplasma test

Identify and freeze

2-10 weeks (According to your requirement)

Notes:        

We offer deliveries for both cell pool and single cell clone.


Dozens of cells, such as HEK293,CHO,HeLa, M14, THP-1, are available.


Amplification is optional if your cell line is to be used for industrial grade productions.


The stability test requires 10 generations of research-grade cell strain and 50 generations of industrial-grade cell strain.


Transfection methods include Electroporation, Nucleofector, Lentiviral based for difficult-to-transfect cell lines.

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