CellMosaic® has designed this personalized antibody-oligo conjugation kit to work with any antibody and an amine-modified oligo. The user supplies the amine-modified oligo, which is readily available from many commercial oligo suppliers. Using the kit components, the customer converts the amine-oligo to a maleimide-oligo (Step 1A in Scheme 1). The customer also supplies their own antibody. Using the kit components, the customer converts some of the surface amines of the antibody to free thiol groups (Step 1B in Scheme 1), followed by reaction of the maleimide-modified oligo to generate the conjugates (Step 2 in Scheme 1). If there are any unreacted thiols of the antibody, they will be subsequently capped (Step 3 in Scheme 1). The one-step filtration type purification step typically provides the resulting antibody-oligo at greater than 90% purity. 

This kit provides materials to perform one (CM53405x1) or three (CM53405x3) antibody oligo conjugations. Scale of the reaction: 0.5−1 mg antibody (protein content).

The kit materials and user manual will be supplied based on your oligo length. Please supply this information when you places the order. Note: If the oligo is over 30 mer, the kit may not be very efficient, please check our custom service for oligo-antibody conjugation. This kit is targeting average 2-4 oligos per antibody. If you are looking for single labeling of antibody, please also check our custom service for oligo-antibody conjugation. 

Requirement for antibody:

• Amount: 0.5−1 mg IgG needed per reaction 
• Preferably >90% pure by gel electrophoresis 

Requirement for amine-oligo:

• Amount: 15−30 nmol oligo content as measured by UV
• Oligo length: <30 mer
• HPLC purified and lyophilized
• Additional components of acid or other molecules containing primary or secondary amine groups and thiol should NOT be present 

Key Features of this Antibody-Oligo Conjugation Kit:

• Offers a simple and easy way to prepare antibody-oligo conjugate
• Loading: 2-4 oligos per antibody
• Easy preparation: <6 h hands on and two day preparation
• All reagents and supplies included for preparation to purification
• Options to choose tailored services at CellMosaic prior to and after conjugation

After conjugation: you can choose to send your sample to CellMosaic for HPLC analysis of the conjugates.

Scheme 1: Synthetic route to antibody−oligo conjugate

Typical Kit Performance Data (LC analysis, CellMosaic)

Antibody information: A therapeutic antibody (human IgG1 subtype)

Oligo information: HPLC-purified 18mer oligo with 5’ NH₂-C6 modification

Kit lot number: S312.S9.122120

Scale of the reaction: 0.5 mg of antibody

Specification of the final conjugates:

Calculated average DOL: 2.46                     Unreacted antibodies: 2.1%

Unreacted oligo: ~5%                                Conjugate Purity: >92% (74% recovery)

Figure 1: Size-exclusion HPLC analysis of the antibody (Ab, red), oligo (blue), and purified antibody-oligo conjugates (green).