Recombinant Protein Expression & Purification

The expression and purification of proteins is one Paragon’s core competencies. With more than 25 years of experience, Paragon has produced and purified hundreds of proteins.

Recombinant Protein Expression

Paragon offers extensive expertise across four expression systems for producing your recombinant proteins, monoclonal antibodies, vaccine antigens or disease biomarkers. Whether for structural studies, functional assays, target validation or high throughput screens, Paragon can help.

Starting from a gene sequence or cDNA, Paragon can clone your gene (with or without affinity tags) into a commercially available expression vector or a custom vector the client provides.

Mammalian

Protein scientists at Paragon have experience and expertise in both transient and stable expression using a multitude of cell lines, including CHO and HEK293 expression systems.

Contracting laboratories have direct access to the scientists working on their projects, and each project is delivered with a specifications sheet and original data on yield, purity, and identity.

Single-use disposable bioreactor technology is extensively used, in scales up to 2x100L actual volume.

Scale

Attached cell lines – T-flasks, roller bottles, cell factories

Suspension cultures

1L & 5L spinners

5L, 14L, 40L, 100L & 200L stir tank bioreactors

20/50L and 100/200L WAVE bioreactors

Cost-effective transient transfection services

Utilizing HEK293 or derivative cell lines, Paragon can deliver high transfection efficiency and protein expression – scalable from 1L to 100L.

Product

Materials to be provided may include cell pellets, concentrated conditioned media, membrane preps or purified protein

Insect

Starting from a DNA sequence, cDNA or recombinant virus, Paragon scientists will clone your gene (± affinity tags) into a commercially available expression vector or a custom vector that you provide.

Cell lines

Sf9 / Sf21 / T ni

Scale

1L, 5L, 10L, 14L, 40L, 200L

Product

Cell paste

Clarified cell culture supernatant

Purified protein

Starting from a DNA sequence, cDNA or recombinant virus

Cloning into commercially available expression vectors (± affinity tags). Veracity of construct demonstrated by restriction digest analysis and sequencing

Co-transfection with AcNPV DNA

Plaque purification of recombinant viruses

Expansion and PCR verification of recombinant viruses

Baculovirus expansion and titer – production of 1L high-titer stock

Time course and MOI study to optimize protein expression

Microbial

Newly-built facilities supporting a vast array of state-of-the-art equipment are leveraged by Paragon’s experienced PhD-level scientists to deliver proteins expressed via microbial systems quickly and economically.

Starting from a gene sequence, cDNA or from almost anywhere a client asks Paragon to begin, our scientists can clone your gene – with or without affinity tags – into a commercially-available expression vector or a custom vector the client provides.

Development

Induction kinetics

Feed strategy

Media optimization

Scale

From Fernbach Flasks to 15L, 50L, 100L, 200L fermentors

Product

Cell Paste

Cell Lysate in a suitable buffer

Clarified culture supernatant

Purified protein

Batch records & specification sheets

Yeast

If you’re looking for protein production via a fed-batch Pichia processes, Paragon has the experienced and credentialed staff to move your project forward.

Our expertise in protein expression spans both P pastoris and S cerevisiae expression systems.

Development

Induction kinetics

Feed strategy

Media optimization

Scale

From Fernbach Flasks to 15L, 50L, 100L, 200L fermentors

Product

Cell Paste

Cell Lysate in a suitable buffer

Clarified culture supernatant

Purified protein

Batch records & specification sheets

Recombinant Protein Purification

Combining skills in protein biochemistry, analytical chemistry and functional assays, Paragon delivers efficient and cost effective solutions in protein purification and process development.

Whether the protein is produced at Paragon or provided by the client as cell pellets, bacterial paste or conditioned medium, Paragon can propose a solution to quickly and cost-effectively purify your proteins to your specifications.

Purification protocols may be obtained from the client, obtained from literature, developed de-novo at Paragon, or adapted from standard protocols available at Paragon. Before your project begins at Paragon, all procedures will be approved by the contracting laboratory.

Final products are accompanied by batch records and specification sheets containing original documentation on yield, purity and identity.

 

Scale

Analytical to Preparative Scale

Microgram to gram quantities

Flow Rates from 1mL/min. to 3L/min.

Chemistries

ION exchange (Q, S)

Affinity (Ni, Co, FLAG, maltose, glutathione, protein A/G)

Gel filtration

Reverse-phase

HIC

Systems

AKTA & Bio-Rad FPLC

Beckman & Waters HPLC

Analytical Methods

SDS-PAGE (Coomasie, Silver)

IEF

Immunoblot

Bradford

ELISA Special High-Performance, High-Dynamic Capacity Resins

Macro-Prep, Fastflow, Hyper D, Poros

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