Custom antibody services for production and purification of modification-specific antibodies.
We offer options for producing phosphospecific antibodies and antibodies specific to other post-translational modifications or activation states of particular proteins as part of Thermo Scientific Pierce Custom Antibody Production Service.
Pierce Custom Antibody Services include powerful capabilities for production and delivery of antibodies that are specific to any protein expression state that can be incorporated into the design and synthesis of the peptide antigen(s) for the protein. These states of protein expression and activation include phosphorylation, acetylation, glycosylation, prenylation, myristolation, ubiquitination, sumoylation, protein cleavage (neo-epitopes), splice variants and isoforms. By the same methods of antigen design and screening-purification, monospecific antibodies can be made to differentiate between particular ligand-binding domains, drug-binding sites, polymorphisms, mutations, species cross-reactivities, and highly conserved proteins.
- Exact monospecificity - produce phosphospecific antibodies or other post-translational modification-specific antibodies or nearly any sort of variant-specific antibody
- Peptide-based accuracy - optimized antigen design and accurate peptide synthesis service maximize antiserum titer and clone production for the intended specific target
- Validated selectivity - reliable methods of positive and negative selection by monoclonal antibody screening or polyclonal antibody purification ensure delivery of antibody with the desired specificity
|Protein Modifications. Selected conditions of protein regulation and variation that can be targeted for monospecific antibody production.|
Details and Additional Information:
Robust methods and processes allow for the development of highly specific antibodies:
- PTM-specific antibodies to quantitate only phosphorylated, acetylated and other post-translationally modified forms
- High-homology antibodies to distinguish between two closely related isoforms or splice variants
- Neo-epitope antibodies to recognize specific protein cleavage sites and conditions
- Binding-specific antibodies to detect proteins only when bound to a drug or ligand
- Fusion protein antibodies to differentiate untagged from tagged proteins (His-tagged, GST, etc.)
The strategy for producing monospecific antibodies is simple and robust:
- Design and synthesize both modified and unmodified peptide forms of the target protein domain.
- Conjugate and immunize with the modified form of the peptide antigen (or whichever form is the desired target).
- Screen and/or purify resulting clones and/or antiserum with respect to both modified and unmodified peptides.
- For monoclonal antibody production, the critical step is careful positive and negative screening of animals (and then hybridomas) to select clones that produce antibodies with the desired specificity.
- For polyclonal antibody production, the critical step is isolation of the specific subpopulation of clones by affinity purification with respect to the unmodified peptide (negative selection) or the modified peptide (positive selection).
Each step in this process is critically important for achieving the desired result. Proper peptide design (and accurate, modification-capable peptide synthesis) ensures that the antigen is effective for antibody production and likely to yield clones with the needed monospecificity. The manner in which carrier protein conjugation is performed determines how well the immunogen elicits an immune response and how effectively the key portion of the peptide is presented as an antigen for production of antibodies that function in the intended assay method(s). Finally, screening and/or purification provides the necessary final identification, isolation, and validation of the specific antibodies.
Custom Phosphospecific Antibody Production
Phosphospecific antibodies (i.e., antibodies for detection and quantitation of phosphorylated proteins of interest) are the most common needs of researchers requesting custom monospecific antibody production. No doubt, this popularity is the result of the essential role of protein phosphorylation in cell signaling, which continues to be the focus of investigation in many areas related to cell biology. The ability to distinguish between different phosphorylation sites within a protein can be quite difficult and complex but is important to understanding cascades and overall protein activity. We specialize in making site-specific antibodies to discrete phosphorylation sites by taking advantage of proprietary antigen design tools (the Antigen Profiler System) and conjugation methods (Targeted Antigen Display Technology). These strategies enhance antigen presentation of phosphopeptides to increase antibody titer and generate more adaptive populations of polyclonal antibodies (i.e., ones that are able to recognize epitopes in different conformations, as found in different assays).