Standard Mouse Monoclonal Antibody Production Service

Standard Mouse mAb Production Service Introduction

Mouse monoclonal antibody has many features such as targeting to a single epitope, high specificity, stable passage, large-scale manufacturing and extensive applications in basic research, medical diagnosis, tumor therapy and radioimmunoimaging. With hybridoma technology, Sino Biological has developed mouse monoclonal antibodies (mAbs), which have advantages of high sensitivity and specificity. We can also provide flexible customized solutions according to customer requirements, aiming to provide customers with high-quality antibodies.

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Standard Mouse mAb Production Service Process

Service procedures Specification Timeline Deliverables Guarantee Price

Antigen preparationQuote!
Please refer to antigen production service

Antigen validation
• Analysis of client's antigen by SDS-PAGE and UV 1-2 days • 2-5 strains of ELISA positive clones
(2 tubes of cryopreserved cells)
• 2-5mL supernatant per clone
• 1-3mg purified antibody (choose a hybridoma cell)
• Experimental report
ELISA positive for immunogen

Immunization and serum titer test
• Pre-immune bleed
• Five Balb/c mice immunization
• Serum titer test
• Final bleed
8-10 weeks

Fusion and Screening
• Fusion and Screening
• Subcloning and cell expansion
• Cell Banking
4-6 weeks

Antibody production & purification
• Appropriate scale hybridoma cell culture
• Purified by protein A affinity chromatography
2-3 weeks

QC analysis
• Analysis by SDS-PAGE and UV
• ELISA validation
3 days

User-supplied antigen requirements:

Recombinant protein antigen Peptide antigen
Antigen quantities 3-4mg/5 mice • KLH/VLP conjugated peptides 5mg/5 mice
Antigen Size  >10kD   Concentration  >0.5mg/mL 
SDS-Page purity  >90% • OVA/Biotin conjugated peptides  2-3mg
Concentration  >0.5mg/mL    Concentration  >0.1mg/mL 
Formulation  PBS, if not PBS, please inquire first   Formulation PBS, if not PBS, please inquire first

Case Study of Standard Mouse mAb Production Service

Sino Biological Inc. has established a high-throughput mouse monoclonal antibody R&D platform. A set of high-throughput R&D programs for mouse monoclonal antibodies are also established through the optimization of immunization protocols, fusion protocols, screening protocols, and high-throughput antibody production and purification protocols. All of our mouse mAbs are validated by ELISA, WB, IHC, FACS et al. , and are tested by multi-tissue and multi-cell.

a)Multi-cell, multi-tissue validation

  • • Human placenta
    Fig 1. Immunochemical staining of human target D with mouse Mab. The left panel: tissue incubated with primary antibody; The right panel: tissue incubated with the mixture of primary antibody and antigen.
  • • Human breast carcinoma
    Fig 2. Immunochemical staining of human target D with mouse monoclonal antibody.

b)Multi-application verification

  • Fig 1. Confocal immunofluorescence analysis of human target D in MCF7 cells. Positive staining was localized to lysosome membrane.
  • Fig 2. Flow cytometric analysis of human target D on Jurkat cells. The histogram were derived from gated events with the forward and side light-scatter characteristics of intact cells.
  • Fig 3. Lane A: Jurkat Whole Cell Lysate. Anti-target D mouse Mab; goat Anti-mouse IgG H&L (Dylight800).
  • Fig 4. Lane A: Hela Whole Cell Lysate; Lane B: Jurkat Whole Cell Lysate; Lane C: Daudi Whole Cell Lysate. Anti-target D mouse Mab; Dylight 800-labeled antibody to Mouse IgG (H+L).

Standard Mouse mAb Production Service Advantages

  • State-of-the-art recombinant protein expression & purification technology, multiple protein expression & purification platforms, rich experience on about 6000 protein production, high success rate of antigen expression.
  • Professional peptide design software and high-efficiency conjugation methods ensure that the immune success rate of designed peptide is greater than 95%.
  • Customized immunization, screening and detection proposed according to customer's final application requirements can be provided.
  • ELISA, WB, IHC, FACS and other validation platforms, multi-cell and multi-tissue verification, aiming to improve antibody screening success rate with low background and high sensitivity.
  • Quoting by steps, charging by nodes, reducing customer's risk and saving costs.
  • Optimized electrofusion technology has more than 3-5 times efficiency compared with PEG mediated cell fusion.

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