Monoclonal Antibodies from mouse and rat

Custom monoclonal antibody development from mice or rats with any kind of antigen. To receive the best results, four different antigen concentrations are used for the immunizations. This ensures an optimal immune response. The spleen from the mouse with the best titer will be used for the fusion.

Step by Step Antibody Development

Davids gives you the opportunity to start or stop with each step. You can for example peform the immunization at Davids and receive the spleen cells or hybridoma to perform the screening in your lab.

Individualization of Antibody Production

The development of monoclonal antibodies is very individual for each customer. We can adapt our protocols for your special needs. This includes immunization protocols, screening conditions or alternative screening substrates. Please indicate at the beginning of the project if you want us to screen with multiple antigens (for positive or negative screening).

Antigen Preparation

Antigen Requirements

Immunization

Immunization Schedules

Fusion

Fusion of Spleen Cells

Screening

Screening & Subcloning

Order

Order Process


Antigen Requirements for our Monoclonal Services

Davids successfully used all kind of antigen for the development of monoclonal antibodies. Customer provided proteins and peptides that represents epitopes of the protein are the most common antigens for monoclonal antibodies. Please have a look at our table to see the requirements for the antigens. It is possible that you first send us your antigen for the immunizations and then send us the antigen needed for the screening. If your antigen is expensive or hard to produce please ask us for efficient immunization and screening to improve the amount of antigen that is needed.

Antigens for the Development of Monoclonal Antibodies
Antigen Amount Instructions Shipping
Protein StructureProtein2 - 3 mg
0.3 - 2 mg/ml
We use your protein for the immunization and Screening-20°C (ice packs) to -80°C (dry-ice) or lyophilized
Protein Expression in E. coliProtein Expression-We produce your antigenYou can ship your expression plasmid at -20°C
PeptidePeptides3 - 5 mg
0.3 - 2 mg/ml
We can synthesize and conjugate your peptides or use your onesLyophilized or with ice packs
Cells for ImmunizationCells106 - 107 cfu / immunizationUse our cell immunization kit to prepare for immunizationsPlease contact us when you want to send cells
Conjugation of Small MoleculesSmall Molecules1 - 10 mg
0.3 - 2 mg/ml
We prepare your small molecules for the immunizationPlease contact us when you want to send small molecules
For Affinity Purification we need additional >0.5 mg protein, 3 mg peptide or 108 cfu

STEP 1: Immunization

2 - 4 months

Titer Development after Immunization We immunize 4 BALB/c mice or 4 black hooded rats with different antigen concentrations to have a broad spectrum of immune responses. We perform ELISA titer determinations to determine the animal with the best response. You are able to receive an aliquot of the test bleed to determine the best titer in your lab as well. With your confirmation we continue with the immunizations or start the fusion and screening.


Monoclonal Antibody Immunization Schedule
STEP Day Description
STEP1: Immunizations
PreimmuneDay 0
1st ImmunizationDay 1
2nd ImmunizationDay 14
3rd ImmunizationDay 28
Test Bleed & ELISA titer determinationDay 35
4th ImmunizationDay 42
5th ImmunizationDay 56
Test Bleed & ELISA titer determinationDay 63Preparation for fusion when titer is good
6th ImmunizationDay 70Optional
7th ImmunizationDay 84Optional
8th ImmunizationDay 98Optional
Test Bleed & ELISA titer determinationDay 112

STEP 2: Spleen Cell Isolation & STEP 3: Fusion

1 day

Monoclonal Antibody Fusion The animal with the best immune response receives a final immunization to boost the antibody producing cells. The spleen cells are isolated for direct fusion with immortal myeloma cell lines. A part of the spleen cells is cryoconserved as a backup. The 3 remaining animals are kept as a backup until the fusion finished.
Alternatively for direct fusion or RNA isolation in our lab, you are able to receive the isolated spleen cells for use in your lab.

STEP 4: Screening for positive clones

2 - 4 months

Monoclonal Antibody Screening Davids grows all fusion cells and screens for positive clones in cell culture plates (alternative screening methods are available). Positive clones are grown in flasks and screened for stability and antigen specificity. Screening against multiple antigens (positive and negative screening) is available. The best clones are isolated and cryoconserved.
You receive small aliquots from purified antibodies from the best 2 - 10 clones for your own applications and to determine which clone is the best for the subcloning. In addition you receive our screening protocol to be up to date and to receive the screening raw data for your protocols. You are able to receive the raw clones before subcloning and perform the subcloning in your lab or to work with the raw clones.

STEP 5: Subcloning

1 - 2 months

Monoclonal Antibody Subcloning We perform subcloning with the limited-dilution method, which ensures that the clone is truly monoclonal and improves the long-term stability. After subcloning you receive all clones frozen or in cell culture for your own lab. The rights of the clones and antibodies that were developed during this project belong to the customer after payment.
Storage of the positive clones for 8 months as a backup is included. Further storage is possible. This enables you to order an antibody production and purification in our lab from your antibody producing clones.

Production & Purification of your Antibodies

1 - 2 months

Monoclonal Antibody Production Antibody production from hybridoma/antibody producing clones that we store for you in our lab or from hybridoma cells you send us. Davids established different purification steps to purify IgG, IgM and other subclasses. For further information go to our Monoclonal Antibody Production page.

Additional Services

We offer multiple additional services for your special needs. For toxic antigens we offer in vitro immunizations or the inclusion of the antigen in liposomes. In addition you are able to receive isolated cells from bone marrow, spleen, lymph nodes and PBMC.
The ELISA raw data are inclusive in our monoclonal immunization service M001.

Additional Services for Monoclonal Antibodies
Cat.No. Description Amount Price
A057Additional Small BleedAnimal DependingPlease ask
A705Additional ELISA Raw Data incl. graphFrom 1 BleedPlease ask
N101Spleen Cell Isolation1 spleenPrice List
N201PBMC Lymphocyte Isolation1 antiserumPrice List
N301Bone Marrow Cell Isolation1 animalPrice List
N401Lymph Node Cell Isolation1 animalPrice List
M006Subclass Determination IgM/IgG1 clonePrice List
M008Storage of cell line for one year1 clone, 3 vialsPrice List
M302Mycoplasm Test (PCR)1 cell culturePrice List

Order Process

You can get your offer for your monoclonal antibodies here: info@davids-bio.com. We keep the order process very simple. You can send us your antigen to:

Davids Biotechnologie GmbH
Roentgenstrasse 3
93055 Regensburg
Germany

The immunizations usually start the same day the antigen arrives. To fasten up the immunization process and help us to connect your antigen with your order ensure you provide the following information:

  1. Your Antigen
  2. Concentration & Buffer Conditions of your Antigen
  3. Our Offer, our order form or your official PO
  4. VAT number (only EU Customers, not German Customers)

If you want to order peptide antibodies just send us your protein sequence and we suggest you peptides that represent your protein. For more information: Peptide Antibodies