Product Description
This routine synthesis service is designed for the small-scale preparation of purified and characterized antibody–oligo conjugates (AOCs), with one oligo per antibody. This single-labeled AOC is especially suitable for diagnostic applications with reliable performance. For routine synthesis of multiple-oligo loaded AOCs, please see RS0003.
The customer provides the antibody to CellMosaic. An amine-modified oligo can be supplied by the customer or purchased by CellMosaic if the sequence is provided. CellMosaic delivers the final conjugate as HPLC purified along with SEC-HPLC analysis data.
Customers can order online by providing oligo information and selecting from various reaction scales, linker types (stable or releasable), and additional optional services.
What's included in the Base Price:
- Labor and materials (excluding antibody and oligo).
- Initial QC and quantitation of the starting antibody by SEC HPLC, and of the oligo by C18 HPLC to check the purity.
- In-process monitoring of the oligo modification and antibody-oligo conjugation.
- HPLC purification to collect the majority of single labeled AOC.
- Final AOC analysis by SEC HPLC to determine purity, aggregation profile, concentration, percentage of unreacted antibody, residual oligo, and multiple AOC species (if separation is sufficient).
- Use of sterilized buffers and plasticware throughout preparation.
Delivery: Final product plus a COA including SEC and AEX HPLC data, concentration, and oligo loading.
Timeline: Approximately 2-3 weeks after receiving the starting antibody and oligo.
Selections and Add-Ons:
1. Reaction Scale: Choose between 1 mg, 3 mg, and 5 mg antibody labeling scale. Typical recovery is 10-30%, depending on antibody and oligo properties. If you do not see the desired scale, please contact us for a custom quote.
2. Oligo Length: Please indicate the number of nucleotides in your oligo so that appropriate handling and conditions can be applied.
3. Linker: Choose from stable maleimide-thiol or releasable VC-PAB linker. if you require a different linker, please contact us for a quote.
4. Antibody Labeling Sites: Choose from surface amines or hinge region thiols. Note: hinge region thiol labeling may results in a higher percentage of unlabeled antibody compared to surface amine chemistry. AEX HPLC purification is recommended for this option.
5. Optional AEX HPLC analysis: AEX-HPLC provides charge-based analysis of AOCs and often offers better resolution. If conjugate peaks are separable, AEX can be used to calculate overall oligo loading. Three samples are generally required for this analysis (oligo, antibody, and AOC). [Click here for more information].
6. Optional 0.2 µm Sterile Filtration: Routine AOC preparation at CellMosaic uses sterilized buffers and pyrogen-free consumables. The final purified product is suitable for in vitro and in vivo studies. However, customers may request additional 0.2 µm sterile filtration after purification (note: this step may slightly reduce the final yield).
Requirements for Antibody:
- IgG Type: Full length IgG (MW of 150KDa), IgM, or F(ab')2 (110KDa). For other antibody fragments or protein oligo conjugation, please contact us for a quote.
- Purity: preferably >90% pure by gel electrophoresis, with minimal aggregation (Lower purity will not prevent AOC preparation but may affect final product quality).
- Amount: The supplied antibody should exceed the selected reaction scale by several hundred micrograms (based on UV protein quantitation). Approximately 20–50 µg of antibody is required for initial QC and optional analyses. If a repeat reaction is necessary, additional antibody must be provided by the customer. Any unused material can be returned upon request.
- Information: Please provide as much antibody information as possible — amount supplied, molecular weight, IgG type, etc. Attach any available QC documentation.
Requirements for Amine Oligo:
- Type: Standard amine DNA oligo with deoxyribose–phosphate backbone. For modified backbones such as phosphorothioate (PS), locked nucleic acids (LNA), RNA, or morpholino oligos, please contact us for a quote.
- Purity: HPLC-purified and lyophilized amine oligo with ≥90% amino-oligo content by C18-HPLC. No additional components containing primary/secondary amines or thiol groups should be present.
- Amount:
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1 mg antibody labeling → >50 nmol oligo
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3 mg antibody labeling → >150 nmol oligo
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5 mg antibody labeling → >250 nmol oligo
(Measured by UV.) Unused materials can be returned upon request.
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- Information: Provide as much detail as possible — amount, molecular weight, sequence, extinction coefficient — and attach QC data from your oligo vendor. Sequence information is required if CellMosaic is to order the oligo.
