Applications
Signature Peptides of Rituximab on a bioZen Peptide XB-C18 Column Using bioZen MagBeads
25617
Separation Mode: Reversed Phase
Reversed Phase
PEPTIDE XB-C18
HPLC
Pharmaceutical/Biopharmaceutical
Therapeutic / Clinical
Biochemical Compound / Nutrient

Applications

Signature Peptides of Rituximab on a bioZen Peptide XB-C18 Column Using bioZen MagBeads
Analytes

Analytes not found

Details
LC Conditions (App ID: 25617)
Column:
Brand Name:
Part No:
Elution Type:
Gradient
Mobile Phase:
A: 0.1% Formic acid in Water
B: 0.1% Formic acid in Acetonitrile
Gradient Profile:
Step No.Time(min)%A%B
108515
24.57525
Flow Rate:
0.3 mL/min
Temperature:
40°C
System:
Agilent Technologies Agilent Technologies
Detection:
LC/MS
Sample Preparation Details
Description:
bioZen MagBeads, Streptavidin Coated - 500mg, Ea
Part Number:
Sample Notes:
1
Dispense
25 µL of bead slurry into a 96 well plate
2
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
3
Add
500µL PBS Buffer to each plate well that contains beads
4
Vortex
bead slurry using mixer or pipette
5
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
6
Add
500µL PBS Buffer to each plate well that contains beads
7
Vortex
bead slurry using mixer or pipette
8
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
9
Add
500µL PBS Buffer to each plate well that contains beads
10
Vortex
bead slurry using mixer or pipette
11
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
12
Reconstitute
beads with PBS buffer to the original slurry volume.
13
Add
10 µL of biontinylated goat anti-human IgG (0.5mg/mL).
14
Transfer
plate for incubation for one hour at room temperature with a shaking speed of 1200 RPM using a deep well plate thermoshaker. Beads should stay suspended during incubation
15
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
16
Add
500µL PBS Buffer to each plate well that contains beads
17
Shake
and mix the bead slurry, using vortex or pipette
18
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
19
Add
500µL PBS Buffer to each plate well that contains beads
20
Shake
and mix the bead slurry, using vortex or pipette
21
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
22
Add
500µL PBS Buffer to each plate well that contains beads
23
Shake
and mix the bead slurry, using vortex or pipette
24
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
25
Reconstitute
beads with PBS buffer to the original slurry volume.
26
Shake
and mix slurry of anti-human IgG coated beads well, using vortex or pipette
27
Transfer
50 µL of bead slurry into each well that contains 25 µL sample. Mix well before transferring
28
Add
cover to plate using polyester plate film
29
Vortex
to mix plate
30
Transfer
plate for incubation for one hour at room temperature with a shaking speed of 1200 RPM using a deep well plate thermoshaker. Beads should stay suspended during incubation
31
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
32
Add
200 µL of PBS Buffer to each well that contains beads
33
Vortex
well and place plate on centrifuge for 2 minutes at 400RPM
34
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
35
Add
200µL of 10mM Ammonium Bicarbonate to each well that contains beads
36
Vortex
well and place plate on centrifuge for 2 minutes at 400RPM
37
Transfer
96 well plate onto magnetic stand for 30 seconds and discard liquid
38
Add
100µL of 0.1% TFA in water to each well and vortex
39
Add
pH paper to check pH value (should be lower than 3)
40
Add
plate cover and shake for 10 minutes at 1200RPM using thermomixer
41
Transfer
plate on centrifuge for 2 minutes at 400RPM
42
Transfer
96 well plate onto magnetic stand for 5 minutes and collect supernatant

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