• Bioproduction Services- High Yield CHO Cells

Bioproduction Services- High Yield CHO Cells

TARGATT™ for Bioproduction of Recombinant Proteins in Chinese Hamster Ovary (CHO) cells. The traditional CHO-cell based protein production is inefficient because they require random insertion and forced amplification of transgenes. ASC’s Master TARGATT™ CHO cell lines enable site-specific, single-copy insertion of large transgenes into a preselected safe harbor locus in the CHO cell genome, for guaranteed gene expression without disruption of internal genes. This site-specific TARGATT™ technology overcomes challenges posed by traditional random insertion methods, and results in high integration efficiency, stable knock-in cell lines, high levels of transgene expression, and reproducibility of clones. The Master TARGATT™ CHO cell lines are ideal antibody and recombinant protein expression.

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Application Notes

Technical Details for Bioproduction in TARGATT™ CHO Cells

Antibody Production in TARGATT™ CHO-H11 cells

Antibody Production in TARGATT™ CHO-H11 cells

Figure 1. Expression of antibodies Ab1 and Ab2 in the H11 genomic hotspot locus of the TARGATT™ CHO-H11 cells yielded ~1g/L of each protein.

Optimizing Docking Site Location in TARGATT™ CHO Cell Lines

Optimizing Docking Site Location in TARGATT™ CHO Cell Lines

Figure 2. Expression of green fluorescent protein (GFP) in various genomic hotspot loci in TARGATT™ CHO cells. TARGATT™ CHO master cell lines were engineered with attP docking site in the well-characterized H11 locus and 4 different proprietary loci, ASC1, ASC2, ASC3, and ASC4. The GFP reporter gene was inserted into each cell line and its expression from each locus was measured using FACS analysis. Reporter expression in the ASC2 locus was 2.5-fold higher than the expression in the H11 locus. The estimated protein yield from the TARGATT™ CHO-ASC2 Master cell line is ~2.5 g/L.

Technical Details

Features

TARGATT™ Site-Specific Insertion

Traditional Random Gene Insertion

Safe Genomic Locus

Yes

No; random

Protein Yield

High

Varies

Gene Insertion Efficiency

High

Low

Reproducibility

Yes; for any protein

No; different from one protein to another

Copy # of Inserted Gene

Single copy

Multiple copies

Stable

Yes

No; Chromosome rearrangement

Gene Silencing

No

Yes

Benefits

  • Genomically well-defined system
  • Stable integration of expression cassette
  • Safe-harbor locus minimizes chances of transgene silencing

Key points:

  • Ablility to achieve > 1 g/l antibody bioproduction using a single site insertion with little optimization
  • Identification of the ASC-X locus has allowed for a further, > 2.5-fold increase in protein expression levels 

Our TARGATT™ technology improves CHO antibody production capability by reducing cost and improving manufacturing feasibility for companies and projects of all sizes. Please let us know if you have questions about your CHO bioproduction project.

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