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Description of Custom Antibody Development Services

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Description of Services

Antibody Formats

Our recombinant antibodies are available in 2 basic formats:

  1. Monovalent Fab antibody, Myc-His-tagged, ELISA positive

  2. Bivalent Fab antibody (Mini-antibody, similar to the F(ab’) 2 format), Myc-His-tagged, ELISA positive.

The molecular weight of our bivalent Mini-antibodies is approximately 110kDa. Each has a small homodimerization domain (termed dHLX, MW = 5kDa), a Myc tag, and a 6xHis tag at the C-terminus of the heavy chain. In some cases, depending on the individual antibody sequence, the purified Mini-antibody may contain fractions of higher molecular weight oligomers. These oligomers do not interfere with standard assays like ELISA, Western Blot, or IHC, and can even be beneficial due to a higher avidity. However if you need to compare affinities or IC50 values of different antibodies, we recommend choosing a monovalent Fab format.

Purified bivalent Mini-antibodies may also contain a small amount of co-purified free dHLX-Myc-His tag, which forms a band running at ~10kDa. This is a well-characterized byproduct of Mini-antibody expression and does not disturb antibody function. Since this co-purifying fragment does not contain any UV-active amino acids (W, Y, F) it will not effect concentration determinations by UV measurement.

Antibodies are also available with other tag combinations or genetically fused to alkaline phosphatase for a slightly higher price. We can also add other fusion partners that can be expressed in bacteria – please inquire antibodies.muc@ab-direct.com for more information.

Production of Antibodies: Your Choice

During the selection procedure we often find several potential antibody candidates, each with a unique antibody sequence. These may have a different specificity and/or a different affinity.

Since each project and customer has individual requirements, we ship the customer small amounts (25-100 µg) of up to 5 unique ELISA-positive antibodies 8 weeks after the start of screening. The customer may test the antibody candidates in specific assays, and select the one that best meets the requirements. We then purify 1 mg of the selected antibody protein for the customer at no extra charge; additional amounts of this or other antibody candidates may be purchased at the same time.

Please note that we cannot guarantee how many antibody candidates will be available for assay, since this is antigen dependent. If we isolate more than 5 antibody candidates against any antigen, they can be purchased for a small charge.

Additional Antibody Material

Bacterial expression clones generated for a specific project are stored for at least 5 years and reserved for exclusive use of the customer unless otherwise agreed in writing. You can order more antibody material at any time. If you choose to order this after delivery of the initial antibody, delivery of the new lot may take up to 4 weeks, and may include an additional set-up fee.

Western Blot-Positive Antibodies

Antibodies that detect native protein may not function in Western-blots, where the protein is denatured. If it is particularly important that your antibody function in Western blots, we can modify the panning procedure to increase the likelihood of generating a Western blot-positive antibody. Although we have a high success rate (>80%) in obtaining Western blot-positive antibodies, we cannot guarantee this functionality. We do offer a selection of micro-purified proxy-candidate antibodies (see above) for the customer to self-select the antibody of choice, or we can perform this service for a small fee.

Solution Panning

Our standard panning procedure screens against immobilized antigens, however sometimes it is beneficial to screen against antigens in solution, for instance if the antibodies should recognize the native protein antigen, and immobilization leads to protein denaturation. In this case, the antigen will be biotinylated before the screening process in order to recover the binding antibodies from solution.

Antigens for biotinylation must be supplied in a buffer that contains no primary amines (e.g. no Tris buffers), and no additives such BSA or HSA. If you are unsure about the suitability of your protein buffer, please contact us.

Antigens: What, how, and when

We can accept a wide range of starting materials as antigens: purified proteins, peptides, peptide sequences, DNA, DNA sequence information, haptens coupled to carrier proteins, and many other compounds. Please let us know if your antigen is not listed here. We will send you an Antigen Data Sheet that you need to fill out in order to give us the necessary information for your project.

Purified Protein Antigens:

The total amount of protein should be at least 0.5 mg, with concentration > 150 µg/ml and purity > 80% as judged by Coomassie-stained reducing SDS-PAGE of a 3 µg sample. The sample buffer must not contain amines (such as TRIS), and should not contain detergents or additives. The buffer used for antibody selection is PBS, pH 7.2.

The antigen will be coated on micro titer plates or immobilized on magnetic beads for the screening process. Please be sure to inform MORPHOSYS if you have any information regarding the immobilization behavior of the protein.

If the antigen is a fusion protein, 0.25mg of the fusion partner is required for library blocking. If the antigen protein is not recombinant, please be sure to indicate the source and provide copies of certificates ensuring that the sample is pathogen-free.

Proteins can be shipped frozen on dry ice or lyophilized.

DNA and DNA Sequences:

If purified protein is not available, we offer our AgX Antigen Expression Service.

Both a DNA sample and its sequence are needed. Please supply the sequence data in electronic format as soon as possible to ensure enough time to choose the best sequence to express for antibody generation. There must be a minimum expressable sequence of at least 240 bases for this approach to be successful.

DNA samples should be delivered as either plasmid or PCR product Please indicate the total amount when sending lyophilized DNA and the concentration if the DNA is in solution.

Peptides and Peptide Sequences:

Peptides used as antigens must be at least 9 amino acids long and must be coupled to 2 different carrier proteins via an N-or C-terminal cysteine residue using 2 different linkers. We use BSA and human transferrin as carriers in our screening process. AbD Serotec offers highly competitive pricing for peptide synthesis and coupling. Any unused peptide will be delivered to the customer with the antibodies. If AbD Serotec organizes peptide synthesis and coupling, the process may take up to 5 weeks before the coupled peptide is ready for use as antigen. If the customer provides the peptide for coupling, it must be at least 4 mg of lyophilized, HPLC-purified peptide, >95% pure, with a cysteine on the N- or C-terminal and no internal cysteines. (If the peptide has internal cysteines, please contact us at antibodies.muc@ab-direct.com to discuss alternate coupling strategies.) The peptide should be water-soluble. Coupling takes up to 2 weeks.

Haptens:

Haptens must be coupled to 2 different carrier proteins using 2 different linkers. We use BSA and human transferrin as carriers for screening. Coupling should be directed and not random. Please contact us for more information.

Rights and Licenses

Antibodies are provided for in-vitro research use only. Please contact us at antibodies.muc@ab-direct.com if you are interested in using the antibodies in commercial applications.

No Antibody = No Charge Guarantee

There is no charge for antibody generation if we fail to isolate at least one ELISA-positive antibody to your antigen. Antigen expression & purification and peptide synthesis are charged separately, and are also dependent on the successful preparation of the antigen. In some cases where the project is highly experimental in nature we request a non-refundable deposit

Invoicing

Invoicing for antigen expression & purification and peptide synthesis occurs once the antigens have been successfully prepared and are ready to enter the screening process. Invoicing for antibody generation occurs when the antibodies are shipped to the customer. Payment is due within 30 days. In cases where there is a non-refundable deposit required, these charges will be invoiced separately at the start of the screening process.

Your Satisfaction is Our Success

We want you to be happy and satisfied with the quality of the products and service we provide and will do whatever we can to ensure that you want to work with us again!