HLA Genotyping

Characterizing and screening for novel and emerging cell therapies.

What is HLA Genotyping and why is it important?

Human leukocyte antigen (HLA) genotype testing has traditionally been used for donor-recipient matching in organ transplantation.  However, HLA genotyping can also be useful for cellular therapies as it can limit recipient response from the patient. This method can also be used for human cell line characterization and testing across biologic manufacturing as it can identify contaminating cells within the human cell line.

Traditional methods such as long range PCR to identify HLA specific alleles are:

Fast and highly scalable
Can suffer from issues leading to ambiguity of results

PathoQuest uses a proven method that eliminates the issues with PCR based NGS sequencing. A hybrid capture step generates the sequencing libraries providing full coverage for all known and novel HLA alleles across 17 loci of the HLA genes.

What we do

Modalities Tested:

  • Allogenic cell therapies
  • Gene modified cell therapies , e.g. CAR-T
  • Stem cell therapies, including mesenchymal stem cells
  • Donor tissue / patient samples
  • Cell lines used across biologic manufacturing
  • Cell bank identity testing

Benefits of NGS for HLA genotyping

Validated method
Faster results
More reliable/robust than long-range PCR
Allele level resolution
Coverage across 17 loci, including all exons
Contamination detection of human cell lines

PCR VS NGS for HLA Testing

WHY THIS APPROACH

Publication: The Importance of HLA Assessment in “Off-the-Shelf” Allogeneic Mesenchymal Stem Cells Based-Therapies

FIND OUT MORE

Publication: Reduced PCR-generated errors from a hybrid capture-based NGS assay for HLA typing

FIND OUT MORE

Publication: Regulatory considerations for developing a phase I investigational new drug application for autologous induced pluripotent stem cells‐based therapy product

FIND OUT MORE

PathoQuest HLA Genotyping Flyer

FIND OUT MORE

Sample requirements

Sample requirements

Shipment & storage Standard turnaround time Fasttrack turnaround time Sensitivity of contamination detection**
Output
Minimum 1×106 cells (or equivalent)

Backup sample required

BSL1 or 2*

Dry Ice /
-80°C
2 weeks 4 days Detection contaminating human cells validated to 5% abundance. HLA Genotype Report

Report of contaminating human cells ≥5%

*Biosafety level classifications can vary between regulatory authorities – contact PathoQuest to discuss.

** NGS should be considered as semi-quantitative in this application.  Abundance figures are provided at the occurrence rate of the contaminating allele as it appears within the read data.  Detection of contaminants may be possible below the validated 5% level.  Please discuss with our experts if this is required.

† Genotyping of genes: HLA-A/-B/-C/-E/-F/-G/-H, DRB1, DRB3/4/5, DQA1, DQB1, DPA1, DPB1, MICA and MICB

*Biosafety level classifications can vary between regulatory authorities – contact PathoQuest to discuss.

** NGS should be considered as semi-quantitative in this application.  Abundance figures are provided at the occurrence rate of the contaminating allele as it appears within the read data.  Detection of contaminants may be possible below the validated 5% level.  Please discuss with our experts if this is required.

† Genotyping of genes: HLA-A/-B/-C/-E/-F/-G/-H, DRB1, DRB3/4/5, DQA1, DQB1, DPA1, DPB1, MICA and MICB

Challenges solved

  • PCR failure due to primer mismatch
  • Balanced amplification across all HLA alleles
  • Base level resolution, including sub population detection
  • Detection of contaminating human cells within a human cell line

“We wished to test for potential contamination in our manufacturing cell line from human cells. Other tests such as CO1 barcoding can detect contamination of non-human cell types but not able to detect human cell contamination of a human cell line. The HLA genotyping assay from PathoQuest was able to fully assure us of the identity of our human cell line.”

OTHER SERVICES

Adventitious Virus Testing

Detection of viral contamination within the manufacturing process and beyond.

READ MORE

Integration Site Analysis

Characterisation of genetic modifications for clone selection, genetic stability and lot release

READ MORE

Identity Confirmation

Genetic characterization of viral and plasmid products for release.

READ MORE

Cell Line Characterization

Biosafety screening and stability testing of manufacturing cells.

READ MORE

In Vivo Replacement

NGS as an ethical alternative to animals in biosafety testing and characterization.

READ MORE

Raw Material Testing

Screening of high-risk inputs such as animal products and media.

READ MORE

Contact us

U.S.

+1 484 212 9360

466 Devon Park Dr
Wayne, PA 19087
United States

Sign up for our latest news

France

+33 (0)1 70 82 17 90

Biopark -Bâtiment B,
11, rue Watt
75013 Paris, France

How can PathoQuest help?

Name(Required)
This field is for validation purposes and should be left unchanged.

U.S.

+1 484 212 9360

466 Devon Park Dr
Wayne, PA 19087
United States

France

+33 (0)1 70 82 17 90

Biopark -Bâtiment B,
11, rue Watt
75013 Paris, France

How can PathoQuest help?

Name(Required)
This field is for validation purposes and should be left unchanged.

Sign up for our latest news