Recombinant protein Expression and Purification

Platform: bacterial mammalian or insect cell lines

Project segmentation to evaluate each milestone reached

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Our standard Protein Expression & Purification stages:

Phase

Time Scale (weeks)

Process

I: Gene synthesis & preparation of target vector 3 Synthesis of protein of interest and cloning into appropriate vector
Choice of purification tag (His-tag/GST-tag) and cleavage site (SUMO,TEV/3C etc.)
Transformation of vector into appropriate host cell type
IIa: Small scale expression 2-3 Small scale (50ml) expression
Optimisation of growth/expression parameters followed by analysis of protein expression by SDS-PAGE, WB etc.
IIb: Medium scale expression 2-3 1 step affinity purification (His-tag/GST-tag) and tag removal (if required)
Analysis of protein purity by SDS-PAGE and WB
III: Large scale expression 3-6 1L scale expression and purification
Typical yield of 1-5mg of purified protein
Further scale will be required to meet your demand

Bacterial:

Phase I

Standard vector: pET
Standard host cell: BL21 DE3

Phase IIa

  • 50ml screens
  • Optimal growth and expression parameters are achieved through the application of a range of different temperature, media & bacterial strains

Mammalian:

Phase I

Standard vector: pcDNA3.1
Standard host cell: HEK 293

Phase IIa

  • Transient expression tests
  • Protein can be expressed intracellularly or secreted into media, depending on protein

Insect:

Phase I

Standard vector: pOET1
Standard host cell: Baculovirus

Phase IIa

  • Amplification of virus stock (100-200ml)
  • Test expression in Sf9 and Tni cells by Western Blot analysis

Phase III

  • Amplification & QPCR titration of virus
  • Purification using His tag

Additional Services

  • Method development
  • Labelling: Selenomethionine, Selenocysteine, N15

  • Validated ELISA assay development
  • Endotoxin removal from protein sample

  • Fragmentation: Fab and Fab’ generation
  • Cell line banking and storage
To request a quote, please fill out the form below and we will get back to you in 1-2 business days. For more information, don’t hesitate to email us at info@bioservuk.com

Protein Expression & Purification Enquiry Form
Clone
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