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Llama antibodies & monoclonal VHH production

VHH production

Eurogentec collaborates with QVQ to produce high affinity monoclonal VHH


Benefits

  • BETTER TARGETING: Due to their small size (~ 15 kDa), VHH can also bind epitopes that are hidden, hence targetting precision is higher compared to normal antibodies;

  • EASIER PRODUCTION: The small size of VHH makes them relatively easy to produce in lower eukaryotes up to very high amounts and purities;

  • UNIQUE PROPERTIES: VHH have a high tissue penetration and are cleared from circulation rapidly. Some VHH can even cross the blood-brain barrier;

  • HIGHER STABILITY: VHH are stable under extreme pH, temperature and against proteases; hence, they keep their native folding and epitope binding capacity under very diverse experimental conditions.;

Besides conventional antibodies, llamas and other camelids also produce single chain antibodies with a unique structure being composed of only two heavy chains and no light chains or CH1 domains.

VHH production

The antigen binding domain of these heavy chain antibodies, VHH, are the smallest naturally occuring antibody binding domains.
Therefore, the generation of such small, single-domain antibody fragments (VHH ~15 kDa) has many advantages over traditional immunoglobulin fragments such as ScFv or Fabs.

VHH production

At Eurogentec, we generate llama polyclonal antibodies targeting multiple antigens, and offer a wide range of additional services. We isolate lymphocyte RNAs from immunized llamas to further proceed with VHH production.

At QVQ, we generate VHH phage display libraries and perform selection and screening of high affinity monoclonal VHH.
Resulting monoclonal VHH can be custom labeled, optimized for “in product stability”, constructed in the desired format and produced on large scale for bulk quantities up to GMP levels.


VHH production

Immunization

Program
Customer dependent; 5 to 10 weeks

Antigen
From the customer or produced at Eurogentec. Protein amount required per immunization: 0.5 mg/ llama (2 llamas are scheduled for immunization).

Standard immunization schedule
Pre-immune bleed (day 0): 4 injections (day 0, 14, 28 and 35), 2 bleeds (day 28 and 42).

Deliverables: the program can be stopped here and you receive pAb (serum or purified Ab).

Testing of immune response
ELISA testing against the antigen is generally performed. This step may require an additional amount of antigen.

RNA extraction

The lymphocytes can be isolated from a blood sample (+/-250ml) followed by RNA extraction. Please note that lymphocytes isolation and RNA extraction must be performed very rapidly after the blood sampling to avoid RNA degradation. RNA is checked for quality prior to sending.

Deliverables: the program can be stopped here and you receive RNA or continued at QVQ with monoclonal VHH selection.

Monoclonal VHH selection

The service of VHH generation includes one or several of the following steps:

Library construction

Deliverables: two libraries with size >107 different clones and >90% insert.

Selection and screening
For high affinity VHH clones + sequence determination.

Deliverables: the sequences of at least 6 binders from 2 families.

Production
VHH single domain antibody production and purification.

Deliverables: 0.5mg protein of the lead clones produced in E.coli and purified (>90%).


Additional services

  • VHH custom labeling for a wide range of applications including immunofluorescence, FACS analysis, in vivo imaging, PET imaging, MRI imaging IF, FACS, IHC and electron microscopy.
  • Large scale production in yeast for animal experiments or crystallography up to GMP production for microdosing studies in human.
  • Optimization of VHH sequence for production, physical and “in product” stability and various types of clinical use.
  • Formatting of the VHH in bivalent, bispecific or custom made formats.

Animal welfare

Animal housing and immunizations are exclusively performed in Belgium. Animal welfare is one of our major concerns and as such, our animal facility fully respects the sternest ethical legislations in force. Our quality management system is ISO 9001 certified and our animal complies with the following association's requirements:

  • Federation of European laboratory animal science associations (FELASA)
  • Uk home office animals scientific procedures act

Each animal is selected with the highest vigilance and identified to guarantee traceability. At any time, experts can visit our facilities and audits can be organized.

More information about our animal facilities.

Confidentiality

Eurogentec and QVQ ensure that your project and your data will be handled under full confidentiality and will never be shared with external parties. Non-disclosure agreements can be executed.

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