What is Custom Antibody Production And What Are The Benefits

What is Custom Antibody Production And What Are The Benefits

Custom antibodies have a variety of applications in research, ranging from therapeutic development to in vitro testing. However, the synthesis of custom antibodies is technically challenging and time-consuming; therefore, choosing a source with a proven track record is crucial. 
 

In this post, you’ll learn several important things to consider while deploying antibodies in research and the benefits that come along with it.  
 

What is Custom Antibody Production? 

The adaptive immune system produces antibodies in response to invading infections. The antibody recognizes the invading bacterium, virus, or other molecules, collectively known as antigens, in a precise lock and key fashion. Hence, antibodies are folded polypeptides or strands of amino acids with antigen recognition sites that identify a specific antigen’s binding site. B-cells of the adaptive immune system create these protein strands. 
 

In biotechnology and sensor development, the specificity of antibody-antigen recognition has several applications. These applications need extremely pure antibody sources. Antibodies are frequently attached to sensor surfaces to detect antigens or used with nanoparticles or other labeled detection platforms to detect antigens. Commercially accessible purified antibodies with antigen specificity are available thanks to custom antibody production. Depending on how they are produced and extracted, these antibodies are classified as polyclonal or monoclonal. Multiple epitopes, or various locations on an antigen, can be recognized by polyclonal antibodies. These types come from a variety of B cell lineages. Monoclonal antibodies are generated from a single B-cell line and identify only one epitope on an antigen. 

The Need For Custom Antibodies 

A custom antibody synthesis might be used when an antibody is required against a specific antigen that is not readily accessible. A 90 percent pure antigen sample is all that is necessary to develop and harvest a custom antibody against that antigen. However, new peptides or proteins can be produced, and antibodies set against an antigen sample are unavailable. In addition, custom antibody production procedures can create both polyclonal and monoclonal antibodies. Consequently, the methods and steps involved vary according to the type of custom antibodies under production, whether monoclonal or polyclonal.  

Critical Applications And Benefits of Custom Antibodies 

Immunocytochemistry (ICC) And Immunohistochemistry (IHC) 

The second most prevalent use for antibodies is immunohistochemistry. Endogenous protein is detected with this approach, which involves probing frozen tissue slices or paraffin-embedded with an antibody. This non-native state can be predicted and imitated by the conjugation procedure used to generate peptide antigens when samples (target protein epitopes) are denatured and crosslinked (fixed) with formalin. 

  • PPAb, RMAb, PMAb are among the recommended options. 
  • If tissue fixation is part of the IHC procedure, use peptide antigens to simulate epitopes’ fixed (crosslinked) state. 
  • With polyclonal antibody production, use peptide antigens to reduce background protein binding. 
  • Polyclonal antibodies are produced for localization, protein identification, and screening. 
  • If more quantitative and extensive analysis is necessary, monoclonal antibodies should come in handy. 
     

Western Blotting (WB) 

The most common use of antibodies, whether custom or not, is Western blotting. Protein samples (typically cell lysates) are separated on a denaturing SDS-PAGE (polyacrylamide gel electrophoresis) gel, transferred to a nitrocellulose or PVDF membrane, and then detected using an antibody. The majority of epitopes of the main sequence are fully exposed for antibody binding because target proteins are denatured and linearized (totally lacking the most high-level structure). 

  • PPAb and RPAb are some recommended alternatives. 
  • If an extracted or recombinant protein (>90 percent pure) is available, use protein antigen.  
  • If pure protein is not easily accessible at a reasonable cost, create and employ a peptide antigen(s) (Typically, expression service and recombinant protein development are not worth the cost in case western blotting is the sole purpose of application) 
  • The manufacturing of polyclonal antibodies is also a good option. (Because monoclonals are too specific to particular conformations of the target, they don’t function as properly as polyclonals when it comes to western blotting.) 

Immunoprecipitation (IP) 

Immunoprecipitation, like ELISA, has a tertiary structural problem. Native proteins from fresh biological samples are captured and temporarily immobilized on agarose beads. On the other hand, immunoprecipitation almost always attempts to capture the entire population of the target protein. (Usually, a later detection step, such as western blotting, is used to identify target proteins’ particular forms or modification states.) As a result, monoclonal antibody synthesis is not commonly necessary. 

  • RPAb, RMAb, PPAb, PMAb are all recommended. 
  • If feasible, employ refined protein antigens (>90 percent pure) or recombinant protein antigens. 
  • If protein is unavailable (or excessively expensive to generate), create and employ a peptide antigen; nevertheless, carefully selecting peptide epitopes is required.  
     

ELISA (Enzyme-linked immunosorbent assay) 

When it comes to quantifying target proteins or particular alteration states of target proteins, ELISA is the preferred method. Target proteins are quantitatively collected from biological samples to microplate wells and then identified using two antibodies. Since ELISA samples are live cell extracts or biological material, it’s critical to employ antigens that preserve and display the target protein’s original tertiary structure. 
 

  • RPAb, RMAb, PPAb, and PMAb are all highly recommended. 
  • For entire, whole-target quantification, recombinant protein antigens are used (i.e., irrespective of post-translational modification state) 
  • Create and employ peptide antigens to distinguish and quantify various target protein forms and modification states. 
  • Monoclonal antibody production is the best option for screening, validating, and obtaining the most consistent and accurate quantitative antibodies, notably for sandwich or competition ELISA with matched pairings. 
     

Conclusion 
 

Custom antibody production is helpful when developing an antibody against a unique or rare antigen. Also, others in the scientific community will benefit from the process, particularly monoclonal antibody production. Once a hybridoma cell line has been created to produce an antibody, it can retain that line for use down the line. Ultimately, whether a monoclonal or polyclonal antibody is required depends on the specific application, project timelines, and if stable cell lines are available. Customers may call on the knowledge and experience of experts in custom antibody production to guide their selections provided you’re dealing with a reputable source. 

Eliza beth

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