ZooMAb^® Recombinant Antibody Technology

• Polyclonal and monoclonal antibody technology and production
• Recombinant antibody technology and production
• Advantages and disadvantages for polyclonal, monoclonal, and recombinant antibodies
• ZooMAb^® recombinant monoclonal antibody technology
• ZooMAb^® recombinant monoclonal antibody unique features

Antibodies have become one of the most important tools in life science research, allowing the detection, quantitation, and determination of changes in proteins and other molecules with respect to time and other perturbations. Our ZooMAb^® recombinant antibodies represent a new generation of monoclonal antibodies that are specifically engineered using our proprietary technologies that provide state-of-the-art consistency and applications performance. ZooMAb^® antibodies are designed with the most user-friendly formulation, handling, and storage features available today and have been validated in multiple immunoassay applications. This platform acquired its unique name because of its proprietary B cell immortalization technology that also allows us to produce monoclonal antibodies in a wide range of species.

Learn how ZooMAb^® antibodies are engineered for consistency using our proprietary recombinant expression system.

Polyclonal & monoclonal antibody technology and production

Polyclonal or monoclonal antibody technology offers different advantages that are desirable for specific applications. Polyclonal antibodies are produced as a result of the activation of numerous different antibody producing B cells in an animal. Hence, they are considered a mixture of antibodies that can react with multiple epitopes on the surface of the antigen. Polyclonal antibodies are generally more tolerant of minor changes in the antigen, such as heterogeneity of glycosylation, polymorphism, or slight denaturation.

With monoclonal antibody production, one B cell produces only one specific sequence of antibody to a given antigen. Hence, a monoclonal antibody is considered a collection of identical antibodies secreted by a single B cell clone with specificity for only one antigenic epitope. In traditional methods, monoclonal antibodies are produced by fusion of B cells with an immortal cell to produce a hybridoma that produces many copies of the exact same antibody. Their specific monovalent nature makes monoclonal antibodies very useful in highly targeted immunoassays and diagnostic applications. However, when compared to polyclonals they are more vulnerable to the loss of epitope through chemical treatment of the antigen.

Recombinant antibody technology and production

In contrast, recombinant antibodies are monoclonal antibodies that are generated by cloning antibody genes into expression vectors and do not involve the use of hybridomas. These antibodies can be cloned from any species using suitable oligonucleotide primers. With this technology, the problems of cell line drift, antibody expression variations, and antibody sequence mutations associated with classical hybridoma production and storage can be avoided. Thus, recombinant antibody technology reduces the use of animals in research and offers exceptional batch-to-batch consistency.

Recombinant antibody production from most species consist of a heavy and a light chain and it is important to note that to accomplish maximum IgG production in mammalian cells, one must achieve a balanced expression of both heavy and light chains. However, expression of these two chains on separate vectors is not an efficient way to generate monoclonal antibody producing cell lines. Use of a single vector carrying two separate expression cassettes provides higher efficiency and both genes are integrated at the same position and expressed independently. For recombinant antibody production, stable cell lines such as the Chinese hamster ovary (CHO) cells, human embryonic kidney 293 (HEK293), or NS0 murine myeloma cells are more commonly employed. CHO cells are most commonly used as a stable expression system and HEK293 cells are highly useful for transient gene expression where protein is harvested within a few days following DNA delivery. These eukaryotic systems offer additional advantages of protein folding, post-translational modifications, and a secretion apparatus that enhances the secretory production of antibodies.

Recombinant antibodies can also be produced in additional formats, such as Fab fragments, single-chain variable region fragments, and as diabodies. Additionally, they can also be expressed in a wide variety of hosts, including bacteria, insect cells, yeast, and mammalian cells. Unlike monoclonal antibodies produced by hybridoma technology, recombinant antibodies maintain a high degree of specificity and low immunogenicity over longer periods of time. Recombinant antibodies serve as highly specific detection probes in research, they are fast finding their niche in the detection of different pathogens and toxins and have great potential in therapeutic applications.

Advantages and Disadvantages of Different Types of Antibodies:
Recombinant, Monoclonals, and Polyclonals

Recombinant	Monoclonals	Polyclonals
Advantages
Increased reproducibility and control Significantly reduced production time Animal-free technology - once antibody sequence is obtained High degree of monovalency Easier isotype conversion Almost no batch to-batch variations Easy to develop humanized version Sequence is known and can always be reconstructed, if necessary	Different clones of antibodies can be generated to different epitopes Hybridoma cells can serve as an infinite source of the same antibody Minimal background and cross-reactivity High homogeneity, consistent and reproducible results High specificity in binding to a single antigen Minimal batch-to-batch variation	Relatively easy to generate and more cost-effective Multiple epitopes on the same protein - more robust signals Better signal with proteins that are expressed in low levels Compatible with broader range of applications Higher flexibility in antigen recognition Useful for detection of denatured proteins
Disadvantages
High degree of technical skills required to develop and express Higher cost to develop and produce, particularly the up-front development and identification of the recombinant antibody sequence	More labor-intensive May be limited in their applications Higher specificity limits their use in multiple species More susceptible to the loss of epitope through chemical treatment of the antigen Hybridoma cell line can drift affecting antibody expression Contamination or loss of cell line Mutation of antibody gene sequence can alter performance	Animal death terminates the source Different bleeds may give different results Immunization of a new animal with the same antigen may lead to different epitopes and different clones may be generated Greater batch-to-batch variability is possible May produce nonspecific antibodies adding to background signal Shared epitopes on different proteins can lead to labeling of proteins other than the antigen protein Difficult to conjugate Requires the use of animals for each production

ZooMAb^® Antibodies: The next generation of recombinant rabbit mAbs

Our first set of ZooMAb^® antibodies are recombinant rabbit monoclonals, but antibodies from several other species will soon be available as well. Compared to commonly used murine species, rabbits have a natural tendency to produce a higher immune response to “difficult” immunogens that include small molecules, peptides, and post-translationally modified proteins. Rabbit antibodies also recognize a greater diversity of epitopes per antigen compared to murine species. Rabbit monoclonal antibodies can detect antigens at the picomolar range with high specificity and provide higher signal intensity in a wide range of applications.

Immunofluorescent analysis of NTERA2 cells was performed using a 1:100 dilution of Cat. No. ZRB5603, Anti-Sox-2, clone 1A2, ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.

Unique Features of ZooMAb^® recombinant monoclonal antibodies

Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them

Formalin Fixed Paraffin Embedded (FFPE) E16.5 mouse embryo sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:200 dilution of Cat. No. ZRB5603, Anti-Sox-2, clone 1A2, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Nuclear staining was observed in subsets of tissue types of developing E16.5 mouse embryo tissue sections.

Lyophilized and pre-aliquoted for maximum stability and convenience

Every ZooMAb^® antibody is provided as a lyophilized product, dispensed in 25 µL equivalences, and stored in a clear polypropylene tube that is specifically designed with an o-ring to ensure an air-tight seal. Each antibody is individually packaged to provide long-term stability, faster shipping, and minimize non-value-added costs that typically incur through cold chain handling of liquids, including the thermal insulated packaging used in transportation. In addition, the lyophilized product is exceptionally stable allowing ambient shipping and storage if needed.

Pre-calibrated concentrations for consistent application-specific titers

ZooMAb^® antibodies are recombinantly produced, purified and pre-aliquoted for you. The material in each tube is always calibrated to work at the titers suggested for the different applications. Every antibody is tested in our own laboratory to work in at least 3 applications before we release them for sale. We ensure that each antibody will perform the way you would expect it to in western blotting, immunofluorescence, and other common applications as listed on our product information page. As these are recombinant antibodies that have been carefully selected, cloned, and purified for high affinity and specificity performance, their effective concentrations are typically lower as compared to conventional antibodies that typically require much more material.

Lysates from mouse embryonic stem cells (Lane 1), Human H9 stem cells (Lane 2), and NTERA2 cells (Lane 3) were probed with Cat. No. ZRB5603-25UL, Anti-Sox-2, clone 1A2, ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Sox-2 (~39 kDa).

Preservative-free for safety and simplicity

Every ZooMAb^® antibody is provided free of biocides like sodium azide or preservatives of animal origin, such as bovine serum albumin (BSA), and finished in a simple phosphate buffered saline (PBS) with trehalose. With ZooMAb^® antibodies, complications arising from potentially hazardous materials or local safety restrictions are minimized and allow its use in both in vivo and in vitro applications.

Multiple package sizes so you never buy more than you need

Every ZooMAb^® antibody is pre-aliquoted at 25 µL lyophilized equivalents, and we offer 4x25 µL packages. Hence, you can use only the quantity you really need for your research and simplify reagent sharing.

The pre-aliquoted design of each pack size also allows you to split your purchase without ever having to resuspend the material, which maximizes product shelf life.

Why you should always look for a ZooMAb^® antibody first

There is never any guessing with ZooMAb^® antibodies. All antibodies are offered at the same pack sizes and price points, no matter what the target is. With ZooMAb^® antibodies you can be sure to get the same feature-loaded product, for every single target, every single lot, stable, pure, and packaged for maximum shelf life stability.

Try a ZooMAb^® antibody today and take advantage of our low entry-level price and pack sizes available for every target we list or with any new ZooMAb^® antibody that we are continuously adding to expand our offerings. Additionally, we would love to have your input on their performance and convenience. You can send us a recommendation through our “Share your Experience” message link. Our ZooMAb^® antibodies are designed and tested with your needs in our mind and are backed by our Antibody Bioguarantee.

It is easy to stay in touch with us to find your target

Use the convenient QR code printed on each ZooMAb^® product label to take you to our website. This home page is where you can find up to date information on all the newest additions to our ZooMAb^® portfolio and can contact us to share your experience with our products.

For Research Use Only. Not for use in diagnostic procedures.

Unless otherwise stated in the Product(s) specifications, any Antibody product is sold for internal research use only and may not be used for any other purpose, which includes but is not limited to, any commercial, diagnostic, or therapeutic use. Our validation processes pertain only to research uses and do not confirm or assure that our antibodies can be used for any unauthorized uses as set forth herein.