When it comes to biotherapeutic protein digestion for peptide mapping or large molecule bioanalysis you’ve got a terribly difficult choice:- Lengthy, cumbersome and error prone in-solution digestion, OR
- Quick, simple and reproducible immobilized enzyme digestion
Proven Thermo Scientific™ SMART Digest™ kits provide a standardized protein digestion routine which is fast, easy and standardized; saving you time and increasing your analytical confidence.
Great right? Absolutely. Still got questions? You’re not alone. So, we’ve summarized some of the top questions posed by our SMART Digest Kit customers and answered them for you. Better yet, you can read them all in the same time that it takes SMART Digest to digest insulin. Here we go, start the clock:
Your FAQs Answered
Question 1: What is a typical digestion time for a therapeutic protein using SMART Digest Kits?
All proteins vary with regards to digestion; you need adjust temperature and incubation time accordingly. Insulins will take approximately 4 minutes and monoclonal antibodies approximately 45 minutes. You can use our Online Digestion Time Calculator to get an idea of how much time you will save.
Question 2: What kind of samples work with SMART Digest Kits?
To date we have successful digestion of all kinds of therapeutic proteins in mouse, monkey, beagle and human plasmas. We also have successful applications in cell lysates, urine and cerebral spinal fluid. If trypsin is able to digest your sample in solution, we’re confident SMART Digest will work too.
Question 3: How much can SMART Digest Kits digest?
We recommend a maximum volume of 200 µL of material. It’ll confidently digest up to 50 µL of plasma (approx. 3.5 mg) and as little as 200 pg if you are really sample limited.
Question 4: Do I have to use a SMART Digest Buffer?
The SMART Digest buffer has a pH 7.2 and is carefully optimized for maximum trypsin activity at elevated temperatures. You can use your own, but it may negatively impact digestion efficiency.
Question 5: Does using SMART Digest at high temperatures give in an increase in protein modifications?
No. In comparison to in-solution digests, a comparable number of PTMs have been observed when screening for deamidation, amidation, methylation and oxidation. No modifications to existing PTMs, such as glycosylated or phosphorylated sites, have been observed. Moreover, the relative amount of Lysine Carbamylation is lower using SMART Digest than when using in-solution digest protocols without urea in the sample preparation.
My question isn’t here.
Don’t worry, it’s very likely your question is among the thirty questions answered in this SMART Digest Kit FAQ document. Download it and start saving time today.
Hopefully that answers your queries. But if it doesn’t, please leave your question in the comments box below and we’ll be sure to get back to you quicker than it takes you to perform an in-solution tryptic digest of an antibody. Boooom!
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Thermo Fisher Scientific
United States
