Monoclonal Antibody Production Service

Monoclonal antibody can be produced via traditional hybridoma technology or recombinant antibody technology. As an internationally recognized antibody expert, Biologics International Corp (BIC) provides monoclonal antibody production services which cover the complete full range of the antibody production process from antigen design through antibody purification. Our dedicated teams strive to work with each customer to successfully develop high performance monoclonal antibodies. BIC ensures that the ELISA titer of the monoclonal antibodies is greater than 1: 128,000. No other company in the world can gurantee the same titer.

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Service Highlights

• No mAb delivery no payment
• Guaranteed ELISA titer (>1:128,000)
• Positive western blot results
• Two positive clones
• In-house antigen design tools
• Soluble antigen production and optimization: maximal native epitopes
• Strict confidentiality: no project-associated information and clones will be disclosed except to the owner

Service Contents

Our monoclonal antibody production service is divided into five phases, enabling each part of the service to be customized. For any question regarding our available packages and how to select these, please contact us and we will be happy to help you.

Phase	Standard Package	Customizable
Antigen preparation (optional)	BIC offers services for peptide synthesis and protein antigen expression for use as the immunogen. Further, we offer antigen design assistance with no additional charge.	Alternatively, customers can provide their own high purity proteins (>90%) for use as the immunogen.
Phase I Immunization (8 weeks)	Five balb/c mice are immunized with multiple rounds of immunogens and test-bled (ELISA test is employed) over a two months period. Serum and ELISA results are then sent to the customer for evaluation.	The number or species of immunized animals is available on request, like two New Zealand white rabbits.
Phase II Cell fusion (3 weeks)	Two mice with the best ELISA titer are chosen, and their spleen cells are harvested for hybridoma fusion. Supernatant from the hybridoma clones will be screened using ELISA, 1 to 2 mL of which is sent to the customer for evaluation.	The number of hybridoma cells or the volume of the supernatant can be customized as per customer requirements.
Phase III Subcloning (5 weeks)	Parental cell lines can be subcloned through limiting dilution to obtain monoclonal cell lines with clonality and long-term stability. This process includes two rounds of limiting-dilution subcloning and screening.	Additional rounds of subcloning can be added at the customer’s request.
Phase IV Production & purification (3 weeks)	Target monoclonal antibodies are produced in cell culture in roller bottles. The antibodies in supernatants are purified via affinity purification using Protein A or Protein G resin.
Phase V QC (1 week)	In the standard package, BIC offers 2-5 positive cell lines, 2-5 mg purified antibody and 100 μg soluble antigen. We guarantee a positive western blot and an ELISA titer of >1:128,000.	We can also offer small scale (up to 500 mg) and large scale (starting from 10 g upwards) antibody production according to customer’s specific needs.

The Process of Monoclonal Antibody Production

After having performed thousands of projects, we have the capacity of producing antibodies in either research quantities or in large scale production, and provide comprehensive service packages for monoclonal antibody development. Contact us for project quotations and more detailed information. Learn more about monoclonal antibody production.

Based on our customized solutions and extensive experience, we provide the following two optional services:

Optional Services

How to Select a Species

Species	Advantages	Disadvantages
Mouse	Common host species for monoclonal antibody generation Small amount of antigen required Easy initial assays Low cost	Not suitable for short peptide antigens Small amount of serum
Rabbit	Good host species for monoclonal and polyclonal antibody production High response to peptide and most protein antigens Broad epitope recognition Moderate cost	Not ideal for highly conserved mammalian proteins
Rat	Small amount of antigen required Ideal host for mouse antigen Easy initial assays Good for small-volume polyclonal antibody projects Low cost	Small amount of serum
Goat	Good for large volume polyclonal antibodies Large volume antibody production Large mammalian host species Easy initial assays	Not suitable for modified peptides Large antigen quantities High cost
Chicken	High specificity against antigen Suitable for polyclonal antibody production Good yield Moderate cost	Difficult to isolate antibodies Not ideal for the most peptide antigens

Monoclonal Antibody Production

Antigen Design

Monoclonal antibody generation begins with the creation of a robust immune response, which requires an ideal antigen. BiologicsCorp's high quality antigen design services include complimentary antigen design assistance for peptide / protein antigens. Based on our sequence analysis techniques and extensive exprience, we will guarantee an immune response against the peptide / protein antigen and deliver 100 μg soluble protein antigen / three peptides (1 mg each).

Cell Fusion

Cell cultured myeloma cells are fused with mammalian spleen cells using polyethylene glycol (PEG) or an electrical pulse that disrupts the cell membrane and allows the merging of two adjacent cells. The fusion results in generating two different types of cells, one with the potential for immortality and the other with the ability to produce a large number of monoclonal antibodies.

Subcloning

Because the population of cells which survive selection remains heterogeneous, subcloning is crucial to ensure that cells are truly monoclonal. Single cells which have the same confirmed isotype are required to assure clonality and achieved through multiple rounds of limiting dilutions.

Production & Purification

After screening assays, monoclonal antibodies are produced by selected hybridoma cells in roller bottles. Protein A, Protein G, and Protein L are three bacterial proteins that can bind to antibodies at sites other than the antigen-binding domain. Using one-step Protein A/G affinity chromatography, high purity monoclonal antibodies can be obtained.

Recombinant Monoclonal Antibody Production

The recombinant monoclonal antibodies are highly reproducibility, specific, and expandable. They are not only identical in each manufacturing batch, but the species, isotype, subtype of a recombinant antibody can also be reformatted. We offer the following two different recombinant monoclonal antibody production services:

• After sequencing the hybridoma cell line, recombinant monoclonal antibodies with a known protein sequence, could be generated within a duration of as less as 7 weeks.
• Using our well-established and highly diverse antibody library, we can generate highly specific, high affinity antibodies in E. coli or mammalian cells in 9 weeks.

The processes are performed in vitro, with no immunization of animals is involved. With years of experiences, we guarantee the quantity and the quality of the antibody.

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