Immunochemistry Services
Overview:
For characterization and preparation of custom antibodies for use in assays, Pacific Immunology Corp. offers the following immunochemistry services:
Affinity Purification:
Affinity purification permits isolation of antibodies against the immunizing antigen, thereby eliminating cross-reactivity that would be present in unpurified serum.
To achieve this specificity, pure antigen (typically peptide) is covalently coupled to an affinity column. Serum is then passed through the column, thereby binding antigen-specific antibodies and allowing non-specific antibodies to pass through.
As a result, Monospecific antibodies, which have the same homogeneity and antigen-binding specificity as monoclonal antibodies, can be quickly isolated from a complex mixture of polyclonal antibodies, providing significant cost and time-savings in comparison to monoclonal projects.
Affinity purification of 25 ml of serum is the most popular option and will typically provide 2-3 mg of peptide-specific antibody. For applications requiring larger volumes of purified antibodies, Pacific Immunology also offers affinity purification of 200 ml of serum using a larger column.
With either purification option, the affinity column is included so that the client can run additional purifications as needed. Most columns can be reused at least 10 times within a 6-12 month period.
- Affinity Purification – 25 ml serum: $450.00
- Affinity Purification – 200 ml serum: $850.00
- Affinity Purification – 25 ml egg yolk: $600.00
IgG Purification:
IgG purification provides an efficient method to separate immunoglobulin (antibodies) from other proteins and molecules in the serum mixture, thereby reducing some background in assays.
Protein G, which has a natural affinity for the constant region of the antibody's heavy chain, is typically used to purify immunoglobulin from rabbit and goat/sheep serum.
However, please note that this will not provide specificity to the immunizing antigen and so affinity purification is the preferred technique for most applications.
- IgG Purification – 25 ml serum: $250.00
IgY Purification:
With custom chicken antibodies, IgY (an avian version of IgG) is collected from the egg yolk. Unfortunately, the yolk cannot be used directly in assays, so IgY Purification is required if an affinity purification isn’t being run. Because of the different structure of the IgY molecule, Protein L is typically used for IgY purifications.
- IgY Purification – 25 ml egg yolk: $400.00
ELISA:
Pacific Immunology offers ELISA services to measure antibody titers against the immunizing antigen. The antigen is bound to the surface of a polystyrene well, blocked to reduce non specific binding, incubated with various dilutions of the test serum and allowed to react with enzyme linked secondary antibody to the appropriate species being tested. The final stage shows color development due to reaction between the enzyme conjugate and the appropriate substrate. Pacific Immunology recommends running an ELISA on the first production bleed of peptide projects before proceeding with affinity purifications.
- ELISA: $50.00 per bleed (covers two rabbits/chickens or 1 goat)