Rabbit Monoclonal Antibody Production Service

Rabbit Mab Production Service Introduction

Rabbit mAb usually has higher affinity and specificity than mouse mAb, it can recognize more novel epitopes and mouse antigens, and plays a key role in the development of therapeutic drug candidates and diagnostic kits. Sino Biological develops rabbit mAb by constructing antibody library to directly obtain antibody genes and their sequences, which is more suitable for the long-term preservation of excellent antibodies and their recombinant expression.
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Rabbit Mab Production Service Process

Service procedures Specification Timeline Deliverables Guarantee Price

Antigen preparationQuote!
Please refer to antigen production service Please fill in the form and send it to
cro-service@sinobiological.com

Antigen validation
• Analysis of client's antigen by SDS-PAGE and UV 1-2 days • Antibody H and L chains sequences
• Purified mAb
• Vectors containing H and L chain, respectively
• Experimental report
ELISA positive for immunogen

Immunization and serum titer test
• Pre-immune bleed
• 2 rabbits immunization
• Serum titer test
• Final bleed
8-10 weeks

Library construction & screening
  4-6 weeks

Antibody production & purification
• Vector construction
• Transient transfection of HEK293 cells
• 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 2.5-4mg/rabbit  • KLH/VLP conjugated peptides 3-5mg/rabbit 
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 Presentation of Rabbit Mab Production Service

Sino Biological Inc. has established a high-throughput rabbit monoclonal antibody development & production technology platform. Rabbit monoclonal antibodies independently developed by this platform have better affinity and antigen detection limit than traditional mouse monoclonal antibodies.

The rabbit Mab developed by Sino Biological Inc. can be validated by multiple assay platforms, such as ELISA, WB, IHC, FACS, et al., and can be tested by multi-tissue and multi-cell. All those verification aims to improve the screening success rate of low background and high sensitivity antibodies.

a)Multi-cell, multi-tissue validation

  • • Human stomach
    Fig 1. Immunochemical staining of human target E with rabbit Mab. The image showing membrane staining of epithelium cell.
  • • Human rectal cancer
    Fig 2. Immunochemical staining of human target E with rabbit Mab. The image showing membrane staining of epithelium cell in intestinal gland.
  • • Human placenta
    Fig 3. Immunochemical staining of human target E with rabbit Mab. The image showing positive staining of trophoblast.
  • • Human liver
    Fig 4. Immunochemical staining of human target E with rabbit Mab. The image showing membrane staining of hepatocyte.
  • • Human gastric cancer
    Fig 5. Immunochemical staining of human target E with rabbit Mab. The image showing membrane staining of epithelium cell.
     
     
  • • Human esophagus
    Fig 6. Immunochemical staining of human target E with rabbit Mab. The image showing membrane staining of squamous epithelium cell. The left panel: tissue incubated with primary antibody; The right panel: tissue incubated with the mixture of primary antibody and antigen.

b)Multi-application verification

  • Fig 1. Immunofluorescence staining of target E in monkey stomach with rabbit Mab. The image showing membrane staining of gastric gland cell. The right panel: merge with DAPI.
  • Fig 2. Immunofluorescence staining of human target E in HeLa cells with rabbit Mab. The image showing membrane staining of HeLa cells.
  • Fig 3. Lane A: A431 Whole Cell Lysate. Anti-target E rabbit Mab; Goat Anti-Rabbit IgG H&L (Dylight800).
  • Fig 4. Lane A: K562 Whole Cell Lysate. Anti-target E rabbit Mab; Clean-Blotô IP Detection Reagent (HRP).
  • Fig 5. Analysis of anti-target E reactivity on HeLa cells.
     

a)Multi-cell, multi-tissue validation

  • • Human mammary gland
    Fig 1. Immunochemical staining of human target F with rabbit Mab. Positive staining was localized to membrane of alveolus epithelium.
  • • Human colon carcinoma
    Fig 2. Immunochemical staining of human target F with rabbit Mab. Positive staining was localized to membrane of colonic gland epithelium.
  • • Human bladder carcinoma
    Fig 3. Immunochemical staining of human target F with rabbit Mab. Positive staining was localized to membrane of transitional epithelium.

b)Multi-application verification

  • Fig 1. Immunofluorescence staining of human target F in SKBR3 cells. Positive staining was localized to plasma membrane.
     
     
     
  • Fig 2. Lane A: MCF-7 Whole Cell Lysate. Anti-target F rabbit Mab; Goat Anti-Rabbit IgG H&L (Dylight800).
     
     
     
     
  • Fig 3. Flow cytometric analysis of anti-target F reactivity on SKBR3 cells. The histogram were derived from the gated events based on light scattering characteristics of viable cells.
     
     
  • Fig 4. Lane A: MCF-7 Whole Cell Lysate; Lane B: A431 Whole Cell Lysate; Lane C: HepG2 Whole Cell Lysate; Lane D: Caco-2 Whole Cell Lysate. Anti-target F rabbit Mab; Dylight 800-labeled antibody to rabbit IgG (H+L).

The Second Generation Rabbit Mab Technology

Secondary generation techniques Liquid and solid phase screening Varied detection platforms can insure the high quality
High affinity
Recognize murine antibodies
High specificity & low background
Easy to be humanized
Identify more novel epitopes
Larger antibody libraries
The second-generation rabbit mAb can be directly modified for genetic engineering

Rabbit Mab Production Service Advantages

  • International leading recombinant protein expression & purification technology, multiple protein expression & purification platforms, rich experience on 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%.
  • Leading antibody library construction and screening technology, reduce the risk of antibody fragment deletions or mismatches and increase the probability of screening for target antibodies.
  • 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.
  • High affinity, 10-100 times more than mouse mAb, in average.
  • One-stop service: From antibody development, vector construction to antibody expression & purification, saving customers time.
  • Quoting by steps , charging by nodes, reducing customer's risk and saving costs.

Contact Us

Tel
+86-10-58628216; +86-10-58628223
Email
cro-service@sinobiological.com