General Production Work Instructions (WI)

6% Casein in 50 mM Borate Buffer

DCN Diagnostics
(7 Day Cured at 37C°)
Doc. No. 10147 Rev. 2 Effective Date: 16JUL2021 Page 1 of 6

Date of Manufacture (DOM): __________________ Prepared by: ___________________

Batch Size: ___________________ Lot #: _________________________ Exp. Date: ___________________

(12 months from DOM)

1.0 INTRODUCTION & SCOPE

This procedure details the preparation of 6% Casein in 50mM Borate Buffer cured at 37°C for 7 Days. The
product is used for blocking and storage buffers as well as other various uses that require casein.

2.0 RESPONSIBILITY
2.1 It is the responsibility of the user to ensure that these work instructions are followed and the
record is complete and accurate.
2.2 The department supervisor will ensure that personnel performing the procedure are trained.

3.0 MATERIALS AND EQUIPMENT REQUIRED

3.1 Materials
Item Vendor Vendor PN LN Expiry Qty. per 1L Qty Used
RMS No.
In-House
diH2O Water,
Water
3.1.1 ≥18.2 MΩ NA NA NA Use Same Day ≤1L
Purification
(diH2O)
System

3.1.2 Sodium Acros AC4194500 1.25g

RMS0146
Tetraborate Organics 10

Sigma- 2.89g
3.1.3 Boric Acid B0252 RMS0031
Aldrich

Casein from Sigma- 60g

3.1.4 C7078 RMS0219
Bovine Milk Aldrich

Sigma- 0.3mL
3.1.5 Proclin300 48912-U RMS0140
Aldrich

Used to
Sodium
3.1.6 DCN 10029 NA adjust pH
Hydroxide, 1N
50mL Conical Various or See 6.13.4
3.1.8 21008-240 RMS0346
Tubes VWR

15mL Conical See 6.13.4

3.1.9 VWR 21008-214 RMS0305
Tubes

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 General Production Work Instructions (WI)
6% Casein in 50 mM Borate Buffer
DCN Diagnostics
(7 Day Cured at 37C°)
Doc. No. 10147 Rev. 2 Effective Date: 16JUL2021 Page 2 of 6

3.2 Equipment
No. Item
3.2.1 Magnetic Stir Bar
3.2.2 Glass bottle or other appropriate container
3.2.3 Class A Graduated Cylinder
3.2.4 Stir Plate
3.2.5 37°C Incubator
3.2.6 pH Meter
3.2.7 Pipette, ID #: ____________, Next cal. Due date: ___________________
3.2.8 Pipette Tips
3.2.9 Deionized Water
3.2.10 Timer, ID #: ____________, Next cal. Due date: ____________________

4.0 DOCUMENTATIONS
No. Item
3002 Operation and Maintenance of pH Meter
3004 Operation and Maintenance of Analytical Balances
3008 Temperature Controlling Procedure
3021 Operation and Maintenance In-House Water Purification System

5.0 SAFETY
5.1 Follow general laboratory safety precautions. Appropriate personal protective equipment should
be worn including lab coat, gloves and safety glasses.

6.0 PROCEDURE
6.1 Perform line clearance prior to preparing reagent. Initial/date: ________/_______________
6.2 Label a bottle with the following information: WIP 6%Casein in 50mM Borate Buffer (7 Day Cured
at 37°C), PN:, Lot #, Date, Expiration Date, Batch Size, and Initials
6.3 Place the bottle on a stir plate and add a stir bar.
6.4 Using a graduated cylinder, measure and add the required amount of diH 2O to the bottle
Item Resistivity Asset Number Cal. Due Date Reference SOP
diH2O, ≥18.2Mꭥ Mꭥ 3021

Item Batch Size Qty Required Qty Added

Qty/1L
X (L) = (L) (L)
diH2O 0.8L

6.4.1 Set the stir plate to medium speed. Initial/date: ________/_______________

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 General Production Work Instructions (WI)
6% Casein in 50 mM Borate Buffer
DCN Diagnostics
(7 Day Cured at 37C°)
Doc. No. 10147 Rev. 2 Effective Date: 16JUL2021 Page 3 of 6

6.5 Measure (+/- 0.05g) and add the Sodium Tetraborate Decahydrate and Boric Acid to the bottle.
Batch Size Qty Required Qty Added
Item Qty/1L
(L) (g) (g)
Sodium Tetraborate X =
1.25g
Decahydrate RMS0146
Boric Acid RMS0031 2.89g

Balance Asset Number#____________ Cal. Due Date: ___________ Reference SOP#3004

Initial/date: ________/_______________
6.6 Verify that the pH is between 7.8 and 8.2.
Record pH: _______________

pH Asset #: ___________ Check the pH logbook to make sure the meter is within calibration.

pH within calibration check box Reference SOP#3002

Initial/date: ________/_______________

6.7 Measure (+/- 0.05g) and add the Casein to the bottles.
Batch Size Qty Required Qty Added
Item Qty/1L
X (L) = (g) (g)
Casein RMS0219 60.0g

Balance Asset #____________ Cal. Due Date: _______________ Reference SOP#3004

6.8 Place bottle on the stir plate, mix it at medium speed for 16-24 hours while stored at room
temperature.
Initial/date: ________/_______________

Time In: ____________ Date in: ____________ Initials: ______________

Time Out: ___________Date out: ___________Initials: ______________

6.9 Adjust the pH with 1N NaOH PN: 10029 as required to reach pH 8.5 (±0.1).
DO NOT USE HCl as this causes casein precipitation.
Starting pH Base Qty Added Final pH
1N NaOH PN: 10029

pH Asset #: ___________ Calibration due date: ________________ Reference SOP#3002

Initial/date: ________/_______________

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 General Production Work Instructions (WI)
6% Casein in 50 mM Borate Buffer
DCN Diagnostics
(7 Day Cured at 37C°)
Doc. No. 10147 Rev. 2 Effective Date: 16JUL2021 Page 4 of 6

6.10 Measure (+/- 0.005mL) and add the Proclin 300 to the bottle.
Batch Size Qty Required Qty Added
Item Qty/1L
(L) (mL) (mL)
X =
Proclin 300 RMS0140 0.3mL

Initial/date: ________/_______________
6.11 Mix solution for at least 30 minutes

Time Start: ____________ Initials: ______________

Time Stop: ____________ Initials: ______________

6.12 Recheck pH and adjust as needed to ensure it is 8.5 (±0.1)

DO NOT USE HCl as this causes casein precipitation.
Starting pH Base Qty Added Final pH
1N NaOH PN: 10029

pH Asset #: ___________ Calibration due date: ________________ Reference SOP#3002

Initial/date: ________/_______________
6.13 Mix solution for 30 minutes

Time Start: ____________ Initials: ______________

Time Stop: ____________ Initials: ______________

6.14 Recheck pH and adjust as needed to ensure it is 8.5 (±0.1)

DO NOT USE HCl as this causes casein precipitation.
Starting pH Base Qty Added Final pH
1N NaOH PN: 10029

pH Asset #: ___________ Calibration due date: ________________ Reference SOP#3002

Initial/date: ________/_______________

6.15 QS to the correct volume based on batch size with diH2O

Item Resistivity Asset Number Cal. Due Date Reference SOP

diH2O, ≥18.2mꭥ mꭥ 3021

6.16 Mix solution for at least 10 minutes at Room Temperature

Time Start: _________ Time Stop: ___________ Initials: ______________

6.17 Place the cap onto the bottle and store in a 37°C Incubator for 7 Days +3hr.
Incubator Asset #: ___________ Calibration due date: _______________ Reference SOP#3008

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 General Production Work Instructions (WI)
6% Casein in 50 mM Borate Buffer
DCN Diagnostics
(7 Day Cured at 37C°)
Doc. No. 10147 Rev. 2 Effective Date: 16JUL2021 Page 5 of 6

Time In: ____________ Date In: ____________ Initials: ______________

Time Out: ___________Date Out: ___________Initials: ______________

6.17.1 After 7-day storage at 37oC, pull the bottle from the incubator and let it acclimate to
room temperature.
Initial/date: ________/_______________
6.17.2 Mix the bottle by inverting several times until the solution is homogeneous.
6.17.3 Allow solution to equilibrate to room temperature then measure the pH of the casein
buffer. Final pH should be between 8.0 and 8.5.
Record pH: _______________

pH Asset #: ___________ Calibration due date: ________________ Reference SOP#3002

Initial/date: ________/_______________

6.17.4 Aliquot Casein Buffer into an appropriate size containers and include ~5mL for QC.
Place vials in the -20° freezer after filling.
Check one: □15mL 10mL max fill □50mL 45mL max fill

# Aliquots:____________ Volume: ________________mL

# Aliquots:____________ Volume: ________________mL
# Aliquots:____________ Volume: ________________mL
Total Volume Aliquoted ________________mL
Initial/date: ________/_______________
7.0 IN PROCESS QUALITY CONTROL
7.1 The below condition must PASS for the product to be released. Mark result with an (X). If FAIL is
checked, initiate a non-conformance report, status label and record the NCMR # below
Condition PASS FAIL
Solution appears translucent clear
Final pH is 8.0 – 8.5

NCMR# or NA: _____________ Initial/date: ________/_________________

8.0 LABELING
8.1 Open the Approved Reagent Label template. Populate the fields with the appropriate information
in the format below, print the labels required plus one, and apply the extra label in the space
provided. Store at the temperature listed below in the WIP area until it is released for use.
 Description: 6% Casein in 50mM Borate Buffer (7 Day Cured at 37C°)
 Part Number (PN): 10147
 Lot Number (LN)

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 General Production Work Instructions (WI)
6% Casein in 50 mM Borate Buffer
DCN Diagnostics
(7 Day Cured at 37C°)
Doc. No. 10147 Rev. 2 Effective Date: 16JUL2021 Page 6 of 6

 Expiry: (12) months from the date of prepared.

 Assign the storage temperature: -20°C.
 DOM/Initial
8.2 Initial and Date on the edge of the label and onto the form.

Affix Label Here FF80HP

9.0 BATCH APPROVAL

9.1 Upon approval, apply the label to the container(s), and move to an approved area for storage.
Comments (Reference rejections, explanations, report numbers, or any other notations):

Performed by: ___________________________________________ Date: ________________________

Reviewed/Released By: ____________________________________ Date: ________________________

REVISION HISTORY
Rev. No. Effective Date Originator Description of Change
Rev. 1 30MAR2021 B. Sheffield Initiation of procedure and document
Updated table in sections 6.9 and 6.12 to match 6.14, Updated 6.10
Rev. 2 16JUL2021 C. Booysen ‘QTY Required’ and ‘QTY Added’ units from L to mL., Removed
duplicated entry sections in 6.12 and 6.14, various formatting.

APPROVALS
Title Name Signature Date

Research Scientist Vaibhav Bajaj Approval on File 16JUL2021

Tech Transfer Scientist Joe Gormican Approval on File 16JUL2021

QS Manager Nichole Navarro Approval on File 16JUL2021

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