We are researching the conjugation of various antibodies to quantum dot microspheres and europium microspheres. The sustainability results have been unsatisfactory. Which stabilizing buffers do you recommend for stabilizing the conjugation step?
Best regards
Hi Ali,
I'd like to store my mAbs in the following buffer for long term:
1. Dilute mAb 1:50 in storage buffer (1x PBS, 1% BSA, 50% glycerol)
2. Prepare the buffer as follows:
- 2.25 mL 100% glycerol
- 2.25 mL PBS
- 45 µg BSA (make 10% solution in PBS)
Vortex thoroughly and store aliquots.
Cheers,
Stöpa
The appropriate stabilizing buffer for conjugating quantum dots (QDs) to antibodies is critical for ensuring the stability and functionality of both the QDs and the antibodies. The buffer utilized is determined by a number of factors, including the type of QDs, the conjugation chemistry, and the antibodies used. However, some general rules can help you choose an adequate buffer:
Example Buffer Composition
A commonly used stabilizing buffer for conjugation might look like this:
- 10 mM PBS, pH 7.4
- 0.1% BSA
- 0.05% Tween 20
Conjugation Chemistry Considerations
- Carbodiimide Chemistry: Often used for conjugating carboxyl-functionalized QDs to amine groups on antibodies. Buffers like MES (2-(N-morpholino)ethanesulfonic acid) at pH 5.0 to 6.0 are commonly used for the activation step, followed by conjugation in PBS.
- Maleimide Chemistry: Used for thiol groups on antibodies. Buffers should be free of primary amines and free thiols.
Final Tips
- Buffer Exchange: Ensure that the antibodies and QDs are in the same buffer system before starting the conjugation to avoid precipitation or aggregation.
- Optimization: Perform small-scale trials to optimize the buffer conditions for your specific QD and antibody combination.
- By carefully selecting and tailoring the stabilizing buffer, you can successfully attach quantum dots to antibodies, assuring the stability and usefulness of the bioconjugates that result.
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