How Much Does It Cost To Make A Custom Antibody?
With advancements in technology, the process of making an antibody has been simplified, and it is now possible to produce antibodies on a large scale. Custom antibody production has been a major concern for most biochemical researchers. However, most people fail to understand the cost of the entire process and prefer to buy pre-produced antibodies for research, medicine, and vaccine production. Most researchers are torn between buying an existing antibody and producing their own. However, both of these methods have their pros and cons. This article will look at how much it costs to produce an antibody.
What Is A Custom Antibody?
Before looking at the cost of producing a custom antibody, we ought to understand what an antibody is and the process of making one. An antibody is a protein component responsible for fighting diseases and providing immunity to living organisms. Once the body experiences a foreign invasion, antibodies will immediately respond. On the other hand, custom antibodies are produced in the laboratory using antigens. A laboratory animal is immunized using a specific antigen, and the synthesized antibody is then collected from the serum. The process of using antigens to induce antibodies in an immunized animal's serum is known as immunization.
What Is The Process Of Producing Custom Antibodies?
To calculate the cost of producing a custom antibody, one must consider the entire process and the requirements. Producing custom antibodies can be easy when using the right technology. It is important to note that there are two types of antibodies, and each is produced differently. The two types are:
Polyclonal antibodies: To produce polyclonal antibodies, animals are injected with specific antigens; common hosts include goats or rabbits. To understand more about this process, we have to examine how to make polyclonal antibodies. Once the appropriate animal is selected, it is immunized with specific antigens, which induce the production of the required antibodies.
Monoclonal antibodies: They are made in vitro through tissue culture methods, unlike polyclonal antibodies, which are produced in live animals. Animals such as mice are used to develop custom monoclonal antibodies, where they are immunized repeatedly. The immunized animal produces specific antibodies within B cells located in the spleen. These cells are then harvested and fused with myeloma cells to create hybridomas. At the same time, the unfused cells, such as excess B cells and Myeloma cells, will die.
The hybridomas, which can be created indefinitely in culture while secreting antibodies, are screened for the needed monoclonal antibodies. This process is often followed by monoclonal antibody sequencing to preserve the genetic code. Those cells that produce the appropriate monoclonal antibodies are cultivated in tissue culture, and the monoclonal antibodies are separated from the medium regularly.
Between the two options, the production of monoclonal antibodies is more expensive and time-consuming. The key difference between the two is that polyclonal antibodies are extracted directly from the animal's serum once it is immunized and has produced an adequate titer of the needed antibodies. On the other hand, monoclonal antibodies must be produced by spleen cells that are then combined with immortal myeloma cells to create hybridomas.
You have to identify amino acids in antibodies through antibody sequencing services during the antibody production process. For antibody production, the following process is used:
- Peptide synthesis (or antigen preparation)
- Selection of the appropriate animal
- Immunization (injection) of the animal
- Collection of the antibodies (serum or B cells)
- Purification process
Each step in antibody production carries its own requirement and specific cost. The costs of each step are then combined to generate the final price of producing an antibody. Let's look at each step in more detail.
Peptide Synthesis
Peptides are often used as the antigens to trigger the production of antibodies. During the production process, you must ensure you have synthesized an adequate amount of high-quality peptides. Historically, the peptide synthesis process was difficult, complicated, and yielded very little output. However, the process has been simplified due to technological advancements; it is now possible to produce peptides on a large scale. Custom peptide synthesis involves the formation of peptide bonds between multiple amino acids to create a specific sequence.
Production of peptides with different designs and exceptional biological responses is now possible due to increased scientific knowledge in the biochemistry field. Methods such as solid-phase and solution-phase synthesis are commonly used. Most peptide synthesis companies prefer using the solid-phase method for producing custom peptides because it is faster and easier to automate. Like protein expression, once peptides are produced, they must be purified to eliminate any side products and impurities that might have accumulated during the production process. Typically, High-Performance Liquid Chromatography (HPLC) is used to ensure the peptide reaches the high purity levels required for successful animal immunization.
Selection Of Appropriate Animals
Choosing the right animal for a specific antigen is one of the essential steps in antibody production. The animal you choose will be injected with target antigens to produce antibodies. In this step, you will incur the cost of sourcing both the lab animal and the antigens. If you don't have access to a lab animal facility, you will typically have to pay for the animal and its housing.
Immunization Of The Animal
The selected animal must be injected with specific antigens to induce the production of antibodies. Several factors, such as the animal species, the type of adjuvant used, and the volume of antigen administered, will affect the immunization process. You will be required to immunize the animal several times and observe the immune response over several weeks. The use of adjuvants is critical here, as they help boost the animal's immune system to produce a higher yield of antibodies.
Collection Of Antibody
Once there is a satisfactory antibody titer (immune response) after immunization, you can then collect the produced antibodies from the serum or the hybridoma supernatant. Different species of animals have unique blood volumes, which determine the amount of serum that can be safely collected. The production bleeds, which are higher-volume bleeds, are performed over time according to ethical guidelines. The collected antiserum is first pooled and centrifuged to eliminate cellular debris before undergoing quality assurance testing.
Antibody Purification
Once you have collected the produced antibodies, they must undergo a purification process to eliminate impurities, such as other proteins or antigens to which the animal was exposed apart from the injected target. Antibodies can be purified in a variety of ways, ranging from crude bulk purification to highly specific affinity chromatography. Antibodies from immunized animals are generated in response to particular antigens. When separated from hybridoma supernatant or serum, the purified antibody can be utilized to probe the specific antigen directly in various assays. Monoclonal antibody purification is usually done in the following three steps:
- Capture
- Intermediate purification
- Polishing
Antibody Production In Bacteria
Apart from using immunized animals, bacteria can be used to produce custom antibodies. The E coli expression system is one of the most common hosts for producing recombinant antibodies. This is because these bacteria grow quickly and can be engineered to create new cells much faster compared to mammalian cell systems. In addition, antibodies produced using the E. coli expression system lack glycosylation, which is beneficial for certain research applications where carbohydrate chains might interfere with the antibody's function.
Importance of Protein In Antibody Production
Proteins play an essential role in the production of antibodies. Antibodies are made by B-lymphocytes; these proteins move through the blood and lymph, attaching to their target antigens and eliminating them from circulation. In many scientific and analytical applications, the ability of animal immune systems to manufacture antibodies that adhere to antigens can be exploited to create probes for detecting compounds of interest. Antibodies are especially well-suited for use as probes due to several critical characteristics, including their high specificity. Procedures for antibody synthesis and protein purification services were established in the 1970s and 1980s. Most laboratories and manufacturers that offer protein expression and purification services also offer custom antibody production services.
Custom protein expression has been made easier and more efficient through the use of ELISA Kits produced by specialized manufacturers, which help measure target protein levels with reliability and consistency. ELISA kit manufacturers have adequate knowledge on how to analyze ELISA data, and you can always consult them when you need advice. For custom protein production, you must select the appropriate recombinant expression system to yield an adequate and high-quality protein.
Protein production services are incomplete without gene synthesis. Genes play a vital role in encoding proteins, and custom gene synthesis helps determine the functions of a particular protein during the protein expression process. The balance between a protein's synthetic and degradative biochemical pathways determines how proteins are generated in any cell. As a result, gene expression should be regulated throughout the custom protein synthesis process.
The nucleus regulates gene expression, especially at the transcriptional level in eukaryotes. It's worth noting that only a certain number of genes are expressed in cells at any given moment. Environmental changes frequently impact gene expression, necessitating certain regulatory safeguards. In addition, the availability of nutrients tends to affect gene expression in prokaryotes. There are several gene synthesis companies that offer quality services and are always available for any form of consultation.
Cost of Producing an Antibody
The following factors will determine the cost of producing a custom antibody:
Method Used in Antibody Production and the Purification Process
The cost of antibody production varies depending on the type and method used in the production process. When calculating the cost, the package should include gene synthesis, recombinant expression, antigens, purification, and protein expression services. Most laboratories and manufacturers specializing in custom antibody services approximate the starting price at around $1,000. Producing a monoclonal antibody can be complicated and more expensive, with an average cost ranging from $6,000 to $15,000. On the other hand, polyclonal antibodies are more affordable and can cost around $1,000 to $2,000 to produce using a simple small molecule antigen.
However, you can lower the cost of producing an antibody per milligram by developing your own antibodies. First, you must secure a reliable and consistent supply of the antigens needed for antibody production. Once you have an adequate supply of antigens, you will save on the cost of antigen production. When researchers source protein antigens, they either start with cell cultures, tissues, or body fluids, such as serum, amniotic fluid, or blood, as the raw materials. Secondly, you should ensure you can access a suitable purification method. In most cases, the purification process tends to be the most complicated and expensive stage.
Stages of Production
Making an antibody is typically most costly in the initial stages, and the production cost often decreases over time. As time passes, you can develop reliable antibody production reagents and methods that help you save on upfront costs. For instance, in the beginning, you will be required to synthesize genes, peptides, antigens, proteins, and other essential components, which can be very costly.
However, once you have produced them on a large scale, you can use them repeatedly to produce large quantities of antibodies. Another example is securing a laboratory animal to use for the secretion of antibodies. If you don't have access to an animal facility, you might be required to incur the cost of buying and keeping the animals in a conducive environment. Once you have acquired the animal, you can use it repeatedly (for multiple bleeds), which will save you a great deal in your next antibody production cycle.
Should You Buy Or Produce An Antibody
Most researchers are uncertain about whether to buy an off-the-shelf antibody or produce it themselves. Purchasing an already-made antibody can be cheap and fast, especially in the early stages of research. This is because you will save yourself the hassle of going through the complicated process of producing a custom antibody. However, as much as buying a catalog antibody looks easy and attractive, there are severe disadvantages to using this method. One, you may fail to get the exact antibody you need, and also, you cannot be certain of the process used to produce the antibody; hence, quality and specificity are not assured.
Final Thoughts
Producing an antibody can sometimes look complicated and expensive; however, most of the costs are incurred in the first stages of antibody production. Once you have set a reliable and consistent supply of the basic requirements, antibody production becomes easier and more cost-effective. If you are considering producing an antibody, you need to make the necessary preparations to ensure a smooth and quick process. Ensure you have all the basic requirements for antibody production ready.
If it is your first time producing an antibody, you should seek expert advice from reliable and professional producers who thoroughly understand the process. This will help you avoid making mistakes that can ruin or complicate the production process.