Enlibio,Your biological research partner

Wuhan Enlibio Biotech Co.,Ltd
Monoclonal Antibody Customization

Mouse monoclonal antibody preparation

Mouse monoclonal antibody profile:

Monoclonal antibodies are produced from monoclonal hybridoma cells in which activated B lymphocytes are fused with myeloma cells. Mouse monoclonal antibodies are the most common and most classical monoclonal antibodies. At the same time, mice are also one of the most common model animals in biology and medicine.

Highly specific

· High degree of homogeneity

· Fast, massive, continuous production

· Wide range of applications, covering multiple areas such as testing and treatment

· Preparation programs and tools mature

· Good application prospects

Qidibio's advantages:

· Unique immune methods save valuable time

· New Advantage MagicTM Immune Adjuvant, Induces Faster, More Immune Effects in 21 Days

· High-performance screening platform for positive hybridomas--Omni-HybridomaTM platform

· MagicTM hybridoma growth factor improves the success rate of cell fusion succession and&ndy expression

Our mouse monoclonal antibodies:

· pM-class affinity and high specificity

· Excellent effect on various antigens

· Can identify and distinguish post-transcriptional modifications (PTM)

· Suitable for formaldehyde-fixed immunohistochemistry

· Large-scale preparation of biologically active antibodies

Service specific content:

Monoclonal antibodies can be prepared using small molecules, polypeptides, modified polypeptides, proteins, fusion proteins, modified proteins, etc. as antigens. At the same time, we also provide antigen preparation services, including DNA including DNA synthesis, plasmid construction, protein expression, protein purification, protein modification, peptide synthesis, peptide modification, cross-linking of peptides and carrier proteins, analysis of small molecule structures, and cross-linking with carrier proteins. If necessary, we also provide antibody purification and antibody identification services.

(Involving post-transcriptional modifications generally include: phosphorylation, methylation, acetylation, ubiquitination, etc.)

Mouse monoclonal antibody preparation process:

Animal Immunity (4-6 weeks)

1. Immunize 5 mice and immunize for 2-3 times;

2. Collect 5 mouse serums for ELISA detection.

3. Provide an assessment report.

Hybridoma preparation (4-6 weeks)

After the valence is qualified, after the customer agrees, conduct the fusion experiment:

1. Select spleen cells from mice with the highest serum titer to fuse with mouse myeloma cells. The

2. Limited dilution of fused cells, 96-well plate cloning;

3. HAT screening hybridoma cells;

4. For immunogen detection, positive 1-10 positive cells were detected and expanded to 48-well plates;

5. Pick positive clonal expansion, micro-80 degrees frozen cells [frozen cells can be stored for more than 1 year];

6. Provide customers with 1 ml of supernatant/positive clones for customer testing. The test results will be returned within 20 days. The

Customers select positive clones based on their own ELISA tests and functional test results; discard the remaining clones. If customers need to save the rest of the clones, they need to communicate in advance, and the costs and storage methods should be agreed separately.

Subcloning (1-3 weeks)

According to the customer's request, select positive clones for subcloning:

1. Each clone was subjected to limiting dilution culture in 96-well plates and ELISA screened for positive subclones.

2. Positive clonal expansion, a small amount of -80 degrees frozen cells;

3. Provide 1 ml of hybridoma cell supernatant for testing by the client; feedback should be provided within 20 days. The

A total of no more than 3 positive subclones are included in the package. By default, the best 3 subclones are amplified and expanded to T-25 Flask culture. After the ELISA is confirmed again, the cells are frozen and frozen. Each subclone freezes 3 tubes. cell.

Ascites preparation (optional) (1-3 weeks)

1. Ascites was prepared from the hybridoma cells selected by the client and 2 mice were immunized. Protein A was purified ascites, and the titer was detected by ELISA.

2. Provide purified hybridoma cells, <5mg/clone;

3. Provide experimental report and residual ascites antibodies;

Note: The cell supernatant can also be selected for purification.

Antibody pairing (optional) (2-4 weeks)

1. Several strains of positive monoclonal antibodies were combined and sandwich ELISA experiments were performed;

2. According to the results of sandwich ELISA experiments, paired antibodies were selected;

3. Issue relevant inspection reports to customers.

Functional verification (optional) (1-3 weeks)

According to the customer's requirements for antibodies, ELISA, WB, and other tests can be performed to verify whether the prepared antibodies can be used for this experiment.

Polyclonal antibody customization

Polyclonal antibody preparation process

Phase 1: Immunized animals   (The conventional host animal is rabbit. Please contact us if you need other hosts)

Designing experimental programs, customers have special needs and need to provide detailed experimental requirements (1 working day).

Antigen synthesis and coupling, small molecules/polypeptides/proteins can all be used as antigenic substances (2-3 weeks).

Animals were immunized and immunized three to four times. Elisa detected immediate titers (6-8 weeks).

Elisa detection Using immunogen as an antigen sample, ensure that Elisa detection titer is 1:10000 or more (1 week).

Phase 2: Antibody purification (Customer selects according to experiment needs)

Antigen affinity purification, antigen-coupled Sepharose 4B prepared antigen affinity chromatography column, only capture specific antibodies against the antigen, the purity of the purified antibody product is high to more than 90%, specificity, high sensitivity, can significantly Reducing background signal-to-noise interference is one of the best experimental methods for antibody purification (1-2 weeks).

Protein A/G purification, specific capture of IgG in the sample, cost-effective, especially suitable for the purification of large-scale samples (1-2 weeks).

Saturated ammonium sulfate precipitation purification, isolation of all immunoglobulins, the highest yield, but the specificity and sensitivity is low, only for qualitative analysis (1-2 weeks).

Phase 3: antibody modification (Customers choose according to experiment needs)

Biotin coupling, antibody modification service: The minimum antibody amount should not be less than 2mg, and the charge above 20mg needs to be charged (1-2 weeks).

HRP-labeled antibody modification service: The minimum antibody amount should not be less than 2mg, and the charge above 20mg needs to be charged (1-2 weeks).

FITC labeling Antibody modification service: The minimum antibody amount should not be less than 2mg, and the charge above 20mg needs to be charged (1-2 weeks).

Phase 4: Polyclonal Antibody Order Delivery

Purified or labeled antibodies can be delivered in lyophilized form or delivered in the form of 50% glycerol.

If you have products in this area, please contact Enlibio Biotech market department.

mail: info@enlibio.com   mail: sale@enlibio.com     Tel:+86-027-87610298
mail: alex@enlibio.com   mail: order@enlibio.com  Tel:+86-027-87610297
Add:B105,Building C1,R&D Building,Zone B,C,D,Wuhan National Bio-Industry Base,No.666 Gaoxin Avenue,East Lake New Technology Development Zone, Wuhan, China