Home Immunology Immunization of Alpacas (Lama pacos) with Protein Antigens and Production of Antigen-specific Single Domain Antibodies
Immunology JoVE (Open Access) Citable · DOI

Immunization of Alpacas (Lama pacos) with Protein Antigens and Production of Antigen-specific Single Domain Antibodies

DOI: 10.3791/58471-v
What you'll learn
  • Immunize alpacas with protein antigens and collect immunized blood samples
  • Isolate B lymphocytes and construct single domain antibody libraries
  • Select and identify antigen-specific single domain antibodies from alpaca libraries
Protocol

A method for the production of single domain antibodies from alpacas, including immunization, blood collection, B-cell isolation, and selection is described.

Difficulty
advanced
Total time
~8–12 weeks (immunization schedule + blood collection + library construction)
Model organism
Alpaca (Lama pacos)
Biosafety
BSL-1

Steps

1
Generate antigen-specific alpaca antibodies via immunization

Immunize alpacas with protein antigens using an established vaccination schedule. Collect blood samples from immunized alpacas to obtain antigen-specific antibody-producing B cells.

▶ 01:39
2
Purify lymphocytes for single domain antibody library construction

Isolate lymphocytes from immunized alpaca blood using standard cell separation techniques. Prepare purified B-cell populations for downstream single domain antibody library construction.

▶ 03:02
3
Select antigen-specific single domain antibodies from library

Screen the constructed single domain antibody library against target protein antigens. Identify and isolate clones expressing high-affinity, antigen-specific single domain antibodies.

▶ 03:59
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