Anti-Peptide Antibody Introduction

Peptides are a primary type of antigen utilized for generating antibodies. They serve as a suitable solution when access to the target protein is limited. Peptide antigens, by limiting the potential epitopes, prove to be excellent for detecting post-translational modification (PTM) sites on target proteins. Furthermore, anti-peptide antibodies exhibit high specificity and affinity, making them ideal for clear antigenic targeting. Creative Biolabs offers comprehensive services in the field of anti-peptide antibodies, leveraging our antibody library construction platform, peptide libraries, and phage display platform to support custom antibody production in the anti-peptide antibody field.

Preparation of Peptide Antigens

Chemically synthesized peptide antigens are small molecules, which makes them minimally immunogenic and only capable of inducing a weak immune response in animals. In order to enhance the immunogenicity of the peptide during the production of antibodies using the hybridoma platform, it is often necessary to cross-link the peptide to a carrier protein. Carrier proteins contain many antigenic determinants that stimulate T-helper cells, thereby promoting B-cell responses. Commonly employed peptide carriers include keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA), ovalbumin (OVA), and bovine thyroglobulin (THY). Among these carrier proteins, KLH exhibits higher antigenicity and is the most frequently utilized peptide cross-linked carrier, while BSA is also commonly used as a peptide carrier. However, the use of BSA as a blocking agent in assay tests imposes certain limitations on the application of antibodies produced through this method.

The process of cross-linking a peptide to a carrier protein is a crucial step in designing synthetic peptides. In general, the N-terminal sequence should be cross-linked by the C-terminal amino acids. On the other hand, internal sequences can be cross-linked to either end. Another consideration for cross-linking internal sequences is the acylation or amination of non-conjugated ends, as the original protein molecular sequence will lack charged ends.

Advantages & Disadvantages of Peptide Antigens

Anti-peptide antibodies are primarily generated using short peptide sequences predicted from the natural sequence of target proteins as immunogens. In certain cases, anti-peptide antibodies offer distinct advantages. Firstly, when your intended antibody application is limited to WB, ELISA, or PTM assays, peptide antibodies are an excellent choice. Secondly, anti-peptide antibodies are an ideal solution when your target protein shares a high degree of sequence similarity with other proteins that are challenging or impossible to express and purify. Anti-peptide antibodies restrict the number of potential epitopes. While the limited number of potential epitopes is usually a disadvantage for most antibody projects and applications, it is an exception when generating PTM antibodies. The limited number of epitopes facilitates the discovery of reactive antibodies that specifically target PTM sites.

However, unlike anti-protein antibodies, which can target conformational epitopes, anti-peptide antibodies can only recognize linear epitopes. Although anti-peptide antibodies can be applied to Western blots and ELISA, it is crucial for the linearized proteinsto include epitopes from the linear peptide antigen used for immunization to ensure positive results.

Creative Biolabs aims to share key points and experiences in anti-peptide antibody production and application to help your project run smoothly.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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