Waters Biopharmaceutical Training Kit - Manual (Page 3)

3
[ CARE AND USE MANUAL ]
Biopharmaceutical Training Kit
Section 2. Sample Preparation
a. Sodium Cesium Iodide:
186007517-2 1 x 30 mL
Sodium
Cesium Iodide
2 μg/μL
30 mL
Nalgene
1. Use without dilution for on-board sample fluidics for
calibration and LockMass reference solutions for analyzing
intact proteins for SYNAPT
® G2, G2-S and Xevo ® G2, G2-S Tof,
and Q-Tof instruments.
b. Horse Heart Myoglobin solution (10 pmol/µl):
186006921-1 1 x 15 mL
Horse Heart
Myoglobin
2.5 mg
15 mL Clear
Nalgene
Bottle
1. Using a 5,000 µL pipette, add 15.000 mL of 90:10
water:acetonitrile + 0.1% formic acid to the bottle containing
2.5 mg horse heart myoglobin (186006921-3).
2. Vortex, and sonicate for 10 minutes. Solution concentration is
167 ng/µL (10 pmol/µL).
3. Using a 1,000 µL pipette, add 500 µL of the stock solution into
a 1.5 mL Waters vial.
4. Label the vial “Myoglobin 10 pmol/µL”.
5. Inject 1-10 µL for LC/MS analysis of the intact protein,
depending on the sensitivity of the instrument.
c. Intact mAb Mass Check Standard solution (1 µg/µL):
186006921-2 1 vial
Intact mAb
Mass Check
Standard
1 mg
TruView Max Recovery
with non-slit PTFE/
silicone Septa
(p/n 186005668CV)
1. Using a 200 µL pipette, add 100 µL LC/MS grade or Milli-Q
water directly to the vial.
2. Vortex and sonicate for 5 minutes. Stock is 10 µg/µL.
3. Using a 20 µL pipette, add 20 µL of stock into a Waters
1.5 mL vial.
4. Using a 200 µL pipette, add 180 µL of 90:10
water:acetonitrile + 0.1% formic acid to the vial, vortex.
Concentration is 1 µg/µL.
5. Label the vial “mAb 1 µg/µL”.
6. Inject 5-10 µL for LC/MS analysis of the intact protein,
depending on the sensitivity of the instrument.
d. Glu-fibrinopeptide B solution (200 fmol/µL):
186006921-3 1 vial Glu-fibrinopeptide B 0.5 mg
4 mL Amber
Screw Top Vial
290002010 1
30 mL Nalgene
fludics bottle
(with cap)
Empty
30 mL Nalgene
Fludics Bottle
(with cap)
1. Using a 5,000 µL pipette, add 2,000 µL of LC/MS grade or
Milli-Q water to vial containing 0.5 mg of glufibrinopeptide B.
2. Vortex and sonicate for 5 minutes. Stock is 160 pmol/µL.
3. Using a 200 µL pipette, add 25.0 µL of stock to one of the
empty 30 mL Nalgene bottles provided in the kit.
4. Using a 5,000 µL pipette, add 19.975 mL of 75:25
water:acetonitrile + 0.1% formic acid to the bottle.
5. Vortex and sonicate for 5 minutes. Solution is 200 fmol/µL.
6. Label as “200 fmol/µL GFP in 25% ACN + 0.1% FA”.
e. MassPREP Enolase Digestion (2.5 pmol/µL):
186006921-4 1 vial
MassPREP
Enolase
digest
1 nmol
TruView Max Recovery
with non-slit PTFE/
silicone Septa
(p/n 186005668CV)
1. Take vial of MassPREP™ Enolase Digest and using a 200 µL
pipette, add 200 µL of LC/MS grade water to the vial.
2. Vortex and sonicate for 5 minutes. Stock concentration is
5 pmol/µL.
3. Using a 200 µL pipette, take 100 µL out of the stock solution
and pipette into a Waters Max Recovery Vial (p/n 186000327c)
or equivalent.
4. Using a 200 µL pipette, add 90 µL of 0.2% formic acid to the vial.
5. Using a 20 µL pipette, add 10 µL of LC/MS grade acetonitrile
to the vial.
6. Vortex. Final concentration is 2.5 pmol/µL.
7. Label as “Control Enolase 2.5 pmol/µL”.
8. Inject 5-10 µL for UPLC ® /MS e peptide map analysis, depending
on instrument sensitivity.