Mouse hybridoma screening & sequencing

1. Raw Materials Preparation

● Antigen preparation

CUSABIO has 16 years of experience in recombinant protein expression and can provide one-stop services from gene synthesis, vector construction to protein expression and purification. Based on the traditional five recombinant protein expression systems (E. coli, Yeast, Insect Baculovirus, Mammalian cell and In vitro E.coli systems), CUSABIO has now launched four dominant protein expression platforms for drug target proteins, namely Virus-like Particles (VLPs), Detergent Micelle, Nanodisc and Nanoparticle platforms. More interesting reading related Virus-like Particles, Detergent Micelle, Nanodisc and Nanoparticle platforms>>

Risk-free for Antigen Production. No protein, No charge

Virus-like particles (VLPs) platform
- Transmembrane proteins (TPs)

• Display the natural conformation of multiple TPs with complete biological activity
• Improve immunogenicity and break the body's own immune tolerance
• Higher abundance of target antigen than that in overexpressing cells
• Best target for phage display as its size is 100-150nm
• Can be used for immune/ELISA/SPR/BLI to help lead antibody discovery

Nanodisc Platform
- Transmembrane proteins (TPs)

• Similar to the cell membrane environment, displaying a complete conformation and maintaining biological activity
• Does not contain detergents, suitable for experiments that interfere with detergents
• Can be used for phage screening
• Can be used for immune/ELISA/SPR/BLI/cell experiments

Detergent Micelle Platform
- Transmembrane proteins (TPs)

• Multiple full-length TPs expression, not only limited to ECD epitopes
• Can be accurately quantified, which is different from VLPs and Nanodisc platforms
• Can be applied to immune/ELISA/SPR/BLI

Nanoparticle Platform
- Small molecular weight protein

• Display natural conformation with complete biological activity
• Enhance immune response
• Can be used for phage screening
• Can be used for immune/ELISA/SPR/BLI/cell experiments

● Reference antibody preparation

Reference antibody refers to drug control antibody. It is usually used to detect the antigen or stable cell line to determine whether the antigen or stable cell line is qualified. In addition, It also acts as a control to screen antibody in FACS. The priority order of reference antibody sequence reference is as follows: marketed antibody drugs, clinical stage (Ⅰ-Ⅲ) antibody drugs, preclinical stage antibody drugs, and FACS antibodies.

● Stable cell line construction

Stable cell line refers to a cell line that continuously and stably expresses or interferes with the expression of a specific gene through stable transduction. In the field of antibody development, stable transfected cell lines can be used as immunogens to stimulate specific immune responses in animals, and can target multiple transmembrane proteins. In addition, stable transfected cell lines are widely used in antibody binding and functional screening experiments. CUSABIO can provide stable cell line construction service, and the specific service contents are as follows:

Cell Pool Screening

Monoclonal Screening

3. Clone Screening

CUSABIO has 16 years of rich experience in the development of hybridoma monoclonal antibodies with mature technical routes, stable and reliable product quality and high success rate. The initial positive rate is as high as about 20%-40%, and the positive rate in FACS is about 10%-20%, which can meet the needs of different types of customers.

4. Quality Control of Recombinant Antibody

CUSABIO conducts antibody functional testing based on customer requirements, including antigen ELISA, cell-based FACS, antigen SPR, and antibody Binning assays. The assays of ELISA and SPR mainly evaluate the binding activity and affinity between antigen and correspoding antibody. The assay of FACS is used to verify the binding activity between the screened antibody and the stable cell line. The purpose of Binning assay is to identify whether the screened antibody and reference antibody recognize the same epitope.