Recombinant antibody expression is a fundamental biotechnological procedure that allows the generation of tailor-made antibodies for research and biomedical purposes. Be it in the analysis of biological phenomena, the use as therapeutic antibodies or for diagnostic purposes – recombinant antibodies have shaped many fields of life sciences. This is why evitria has specialized in recombinant antibody expression services, providing partners around the world with custom-made products.
But how is this procedure performed? To provide an answer to this, we will have a more detailed look at the various steps involved in the expression of recombinant antibodies.
Recombinant antibodies, also known as engineered antibodies, are produced through genetic engineering techniques. They involve combining genetic material encoding the heavy and light chains of an antibody to create a functional antibody molecule, binding to a specific epitope.
In contrast to the polyclonal antibody production process, monoclonal antibody production (and also rAbs production) is an ex vivo procedure without the need for live animals. Recombinant protein expression is based on recombinant DNA technology, also involving several other techniques in molecular biology, e.g. post-translational modifications and optimizations.
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These antibodies are used in research, diagnostics, and therapeutics. They offer precise control over antibody characteristics and can be engineered into various formats, such as full-length antibodies or smaller antibody fragments.
Recombinant antibodies provide advantages in terms of specificity, affinity, and effector functions. They have revolutionized antibody engineering and opened up possibilities for developing novel diagnostics, therapeutics, and research tools.
By manipulating nucleic acids and expressing them in host cells, recombinant antibodies can be generated with specific properties. They are used as therapeutic means, e.g. for receptor blocking or the fight against viruses, and also play a crucial role in in vitro studies, allowing for the detection, quantification, and characterization of antigens.
Monoclonal antibodies (mAbs) and recombinant antibodies (rAbs) differ in their production methods and characteristics.
Monoclonal antibodies are immunoglobulins derived from a single clone of plasma cells, resulting in a highly homogeneous population. They are produced by immunizing animals and isolating a single B-cell clone that continuously produces the desired antibody, often involving hybridoma technology.
Recombinant antibodies, on the other hand, are generated through genetic engineering. Genes encoding the variable regions of the heavy chains and light chains of an antibody are cloned and expressed in host cells. This allows for precise control over antibody properties and the production of large quantities.
Recombinant antibodies offer the advantage of producing human-compatible antibodies, reducing the risk of adverse reactions. They also provide flexibility in antibody engineering, allowing for modifications and the generation of bispecific antibodies.
The definition of recombinant antibody expression often refers to the process of producing antibodies using recombinant DNA technology. This involves genomic techniques such as cloning the genes that encode the antibody of interest and then expressing them in a suitable host organism, such as bacteria, yeast, or mammalian cell lines.
By using recombinant antibody expression, it is possible to produce large quantities of a specific antibody in a controlled manner, which can be used for a variety of research, diagnostics, and therapeutic applications including various standard assay types.1
Recombinant antibody expression usually involves the following stages, which can vary depending on the ultimate objective of the project2:
Recombinant antibody expression requires certain conditions: The production of recombinant proteins on a larger scale is a high-tech process which necessitates trained labor and a fully equipped biotech laboratory, as specialized instruments and substances like agarose gel, ampicillin, glutathione imidazole are not to be dealt with in every environment.
Genetic information as well as expression vectors are also needed. Additionally, a cell culture such as prokaryotic microorganisms or mammalian cell lines, and downstream processing and conjugation with small molecules are essential.
The choice of the expression systems for rAbs depends on various factors, such as the size and complexity of the antibody, the desired yield and quality of the final product and available resources.
Bacteria such as Escherichia coli (E. coli), insects or yeast are often used to express rAbs since they are easy to manipulate and have a fast growth rate. However, these three systems may not be able to properly fold and assemble complex proteins, which can lead to low yield and poor product quality.3
Mammalian cell lines, in particular CHO cells or HEK293 cells, are thus the often preferred and most commonly used expression system for recombinant antibody production.4
HEK293 cells (human embryonic kidney cells) are often chosen as hosts for protein expression because of their quick transfection rate, easy handling, and high protein production. Additionally, they are simple to propagate and sustain, and are compatible with various transfection techniques. However, they are restricted to research environments due to various limitations such as their propensity to clump together.
CHO cells (Chinese hamster ovary cells) are vigorous host cells that exhibit strong growth in suspension cultures, adapt effortlessly to serum-free media, and can generate and secrete recombinant antibodies in the multi-gram scale with higher specificity. Due to their origin in hamsters, these cells are less prone to human viral contaminants, yet still possess the capability of performing glycosylation that is compatible with humans.
In CHO cell antibody production, nutrients and supplements act as an inhibitor that stimulates growth and increases enzyme concentration.1
Recombinant antibodies are necessary for various reasons. They offer versatility in antibody formats and engineering, allowing for the generation of tailored antibodies with specific properties.
Antibody fragments, a type of recombinant antibody, have smaller sizes and unique advantages such as improved tissue penetration. They are particularly useful in applications where full-size antibodies may not be optimal.
Recombinant antibody technology enables the production of bispecific antibodies, which can simultaneously bind to two different antigens. These antibodies have promising applications in targeted drug delivery and modulating immune responses.
Proper formation of disulfide bonds, crucial for antibody stability, is ensured through recombinant antibody expression. This improves antibody quality and reduces aggregation.
Furthermore, recombinant antibody systems provide the ability to generate fully human or humanized antibodies, reducing the risk of immunogenicity in therapeutic applications.
At evitria, we are specialized in recombinant antibody expression services based on CHO cells (Chinese Hamster Ovary cells), which allows us the high-throughput production of high quality antibodies with an outstanding speed.
Since we are entirely focused on antibody and protein expression, we can support partners worldwide with specialized knowledge in various questions and challenges along different projects – from a first pilot study to the delivery of different antibody products.
Recombinant protein expression involves cloning genes encoding the protein of interest into host cells, such as bacteria, yeast, or mammalian cell lines. The host cells then transcribe and translate the genes, producing the desired protein. Protein secretion can occur into the supernatant or retained within the cells. Techniques like phage display and fusion proteins are utilized to optimize expression and enhance protein production. The titer and quality of the recombinant protein are determined by factors such as the choice of host cell, transcription efficiency, and post-translational modifications.
Recombinant antibodies are generated by cloning genes encoding the variable regions of the heavy and light chains of an antibody into host cells, commonly mammalian cell lines. The host cells then produce the recombinant antibodies, which can be full-length or engineered into smaller antibody fragments. Recombinant antibodies can be fully human antibodies or humanized to reduce immunogenicity. Once produced, recombinant antibodies bind specifically to their target antigens, such as IgG1 (an IgG subtype), in research, diagnostics, and therapeutics. Their chimeric nature and human compatibility make them valuable tools in biomedical applications.