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Guaranteed Mouse Monoclonal Antibody Production Services

Mouse Monoclonal Antibody Development, Custom Mouse Monoclonal Antibody, Hybridoma Generation

Catalog No. Product Name Size List Price (US$) Quantity
BS010A ELISA Guaranteed Mouse Monoclonal Antibody production 1 package 4500.00
BS010B ELISA Guaranteed Mouse Monoclonal Antibody production + Standard Peptide Synthesis + Peptide Conjugation 1 package 4900.00
BS010D ELISA Guaranteed Mouse Monoclonal Antibody production + DNA Subcloning + Bacterial Expression and Purification of Protein Antigen 1 package 7000.00
Description

The most comprehensive hybridoma-based mouse monoclonal antibody production services:

custom production of monoclonal antibodies by hybridoma technology against non-modified, phospho-specific, methylation-specific, and acetylation-specific peptides and proteins as well as small molecular compounds; for ELISA, Western blot, IHC, and flow cytometry applications.

Hybridoma generation is the oldest and the most cost-effective method for monoclonal antibody development. It has been successfully used with mouse, rat, hamster, rabbit, sheep, and human. Our experience is mostly from mice. The workflow consists of: animal immunization, cell fusion with PEG or electro, screening antibodies with applications of ELISA, flow cytometry, tissue microarray, and such, and then hybridoma subcloning and expansion. It was reported that the electro fusion method gave 10 times of clones out of the PEG-based hybridoma fusion method. Screening of antibodies for various applications may be done by CROs like Syd Labs or clients themselves. After the number of hybridoma clones narrows down, antibody activity is confirmed by clients or in house during antibody screening and hybridoma subcloning.

BS010A: $4,500 for a standard 24-week process of mouse immunization and hybridoma development following the standard protocol of mouse monoclonal antibody development.

Phase I, mouse immunization: Immunization and boosts of 5 Balb/c mice following the standard immunization protocol and ELISA screening of mouse polyclonal sera.
Phase II, spleen fusion: Fusion of splenocytes to mouse myeloma cells and ELISA screening to isolate antigen-positive hybridoma cells from an initial count of approximately 3000 fusion clones. Supernatant (conditioned media) from 10 of the final 2-3 ELISA-positive clone population pools will be sent to clients for evaluation of specific applications.
Phase III, hybridoma development: Subcloning of the ELISA-positive clone population pools by limited dilution to isolate individual hybridoma clones of interest, and expansion of the individual hybridoma cell lines. 

You provide: Ten mg of peptide (purity > 85%, length > 12 amino acids) or 4 mg of soluble protein (concentration > 0.4 mg/ml, purity > 90%, molecular weight > 20 kDa) for immunization, and additional 5 mg for antigen affinity purification.

Deliverable:
* About 200 ul supernatant from 10 ELISA positive hybridoma cells for evaluation;
* 1 positive hybridoma cell line;
* Remaining antigens;
* ELISA results.

The highest quality standard in the field of mouse monoclonal antibody production services: guaranteed ELISA titer > 1:40 000.

The prices of custom mouse monoclonal antibody development services will be adjusted for the following requirements:

a. Peptide (12-15 amino acids, >85%) design and synthesis +$250 for standard, +$450 for mono-phosphorylated, +$650 for di-phosphorylated, +$700 for mono-methylated (K or R site), +$650 for di-methylated (K or R site), +$700 for tri-methylated (K site), +$400 for acetylated;
b. Protein: DNA subcloning +$350, bacterial expression and purification +$2,200;
c. Additional boost for higher ELISA titer of the antisera +$150/injection;
d. Additional positive hydridoma cell line +$600.

Questions and answers:

Question: Do you have a lab facility in US for your mouse monoclonal antibody development service?
Answer: Syd Labs has always tried its best to save clients' cost and provide convenience to clients. Syd Labs has provided various antibody services from facilities in China with the lowest prices in the past decade. Syd Labs added labs for monoclonal antibody services in California and Massachusetts to serve local clients in US, including San Francisco, San Diego, Boston, and New York.

Question: What is your advantage of monoclonal antibody production services in US?
Answer: Many US clients prefer to work with CRO companies (Contract Research Organization) in US for various reasons. For example, clients can do animal immunization by themselves or university core facilities, and send the spleen to Syd Labs for hybridoma fusion and later subcloning and expansion. Polyclones from hybridoma fusion should not be frozen before subcloning because it is easy to lose the monoclone to pursue during the process. The cells must keep growing during screening and activity confirmation. If a client prefer to screen antibodies by themselves with their own functional assays such as flow cytometry, the turnaround time from supernatant shipping to screening results back has to be no longer than 4 days. For such a collaboration process of custom monoclonal antibody development, it is better to have a domestic CRO like Syd Labs.

Question: Can you do DNA immunization for monoclonal antibody development?
Answer: Yes, Syd Labs does monoclonal antibody production with immunization from DNA. Please request quote for DNA immunization. Syd Labs can also use peptides, small molecular compounds, proteins, and cells expressing a protein as the antigens. One needs to do KLH or BSA conjugation if the molecular weight of the antigen is smaller than 20 kDa. During mouse immunization for monoclonal antibody development, Syd Labs may make a combination of various immunogens such as immunization with DNA or cells and boost with proteins.

Question: Can you do RIMMS for mouse immunization?
Answer: Yes, Syd Labs uses Repetitive IMmunizations Multiple Sites (RIMMS) as the alternative immunization protocol to the standard immunization protocol of mouse monoclonal antibody production. Please request quote for RIMMS immunization. Normally the timeline of RIMMS immunization is shorter than 30 days. Some CRO companies promote their monoclonal antibody services as "60 day monoclonal antibody development" or even 29 day. It is possible but it really depends on what clients or the CRO companies are looking for. For example, one may get more IgM antibodies. Or they do not count the timeline of hybridoma subcloning. One may shorten the timeline of mouse immunization but not the hybridoma fusion, screening, and subcloning.

Question: What are the most important parts in custom monoclonal antibody development?
Answer: No matter if clients develop monoclonal antibodies in house or by CRO, one must have two things: one is a reliable high-throughput assay to evaluate antibody candidates, and the other great results of animal immunization. Depending on clients' antibody application, one needs to develop:  functional assays for therapeutic application, immunoassay with real samples for clinical diagnostics application, and tissue microarray for reagent development.

References:

1. "Mouse monoclonal antibodies 1G, 6A, and 10D were produced for this project by Syd Labs, Inc. (Natick, MA) using full-length recombinant human acetylcholinesterase (rHuAChE) as immunogen. Large quantities of monoclonal antibodies 1G, 6A, and 10D were produced in mouse ascites fluid and purified by Syd Labs, Inc." Immunopurification of Acetylcholinesterase from Red Blood Cells for Detection of Nerve Agent Exposure.

2. "Antibodies were generated in mice by Syd Labs, Inc. using standard hybridoma fusion technology." Monoclonal Antibody That Recognizes Diethoxyphosphotyrosine-Modified Proteins and Peptides Independent of Surrounding Amino Acids.

Syd Labs also provides other monoclonal antibody development services:

Rabbit monoclonal antibody development services
Alpaca/llama monoclonal antibody development services

After great hybridoma clones have been developed, if clients want to file patents, protect the antibodies using the digital format, or develop antibody drugs, clients can ask Syd Labs to clone and sequence the hybridoma antibody variable regions (VH and VL), do transient expression of chimeric antibodies (scFv or complete IgG) in CHO or HEK293 cells, or generate stable cell line expressing antibodies.

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