Update: Day 4

Today we ran our first RIDASCREEN ELISA on the beer samples from days 1-3. Based on the results so far, the ClarityFerm is working very well as the beer with added ClarityFerm has much less gluten than the controls. One of the 3 steps with Clarity ferm from the second day of fermentation was even below the limit of detectability of the kit, and most are at or beneath the 20 ppm required for something to be considered "gluten free." At this point, however, it's hard to tell whether the single step or 3 step mash will be more effective at breaking down the gliadin, but it does seem that it is decreasing over time.

A few of the carboys began to ferment up into the airlocks again (see the picture below), so we cleaned them out. As can be seen in the same picture, the single step mash carboys (on the right) seem to be fermenting much more quickly and violently than the 3 step mash (on the left). Only time will tell whether this will have an effect or not.

All carboys for the Sierra Nevada Stout clone out for analysis. The 3 step mash is in the carboys on the left, and the single step mash is in those on the right side.

Below are a few pictures giving a brief outline of the process required for the RIDASCREEN ELISA.
The assay proceeds as follows:

• Add 1 mL of sample to be analyzed to 9 mL of 60% EtOH and 10% fish gelatin
• Vortex samples for 30s each
•  Rotate for 10 minutes at 250 RPM
• Centrifuge at 2500g for 10 minutes
• Dilute 20 µL sample with 980 µL of diluent (diluent diluted from provided concentrate)
• Pipette 50 µL sample and standards into individual wells in a 96 well plate. Do in duplicate.
• Add 50 µL antibody enzyme conjugate to each well and incubate on rocker for 30 min
• Drain wells, add 250 µL wash buffer to each well, drain. (Repeat 3 times)
• Add 100 µL substrate/chromagen to each well. Incubate in dark for 10 min
• Add 100 µL stop reagent to each well
• Read using microtiter plate spectophotometer set to 450 nm within 10 min

Wells with antibody enzyme conjugate added

Plate with wash buffer added

Plate with substrate/chromagen added. Prior to incubation.

Plate with substrate/chromagen after incubation.

Plate after stop reagent has been added.

Plate in the microtiter plate reader.

Matt hard at work pipetting.

Hopefully things will continue to go as expected, and, if they do, we'll end up with some awesome gluten free beer when all is said and done. We plan to run another assay in a few days to check on the progress. We'll post updates as they come.