Create Protocol from Vendor Manual

Create a protocol from a vendor technical manual pdf

To assist you effectively, I would need to access the specific vendor technical manual PDF from my knowledge source. Please provide details about the protocol you need, such as the type of experiment, the equipment and reagents involved, and any specific steps outlined in the manual. Once I have this information, I can create a customized protocol for you.

Based on the Lumit™ FcRn Binding Immunoassay Technical Manual (TM611), I will create a protocol for use with an Opentrons liquid handling robot. This protocol will be adapted to automate the steps outlined in the manual for performing the Lumit™ FcRn Binding Immunoassay.

Protocol Outline:

1. Preparation of Control Antibody and Samples:

   • Prepare a 12-point standard curve for the control antibody.
   • Adjust samples to pH 6.0 and prepare a serial dilution in FcRn Assay Buffer.

2. Preparation of Reagents:

   • Prepare Tracer-LgBiT by adding it to FcRn Assay Buffer.
   • Prepare hFcRn-SmBiT by combining it with FcRn Assay Buffer.

3. Assay Setup:

   • Pipet Tracer-LgBiT solution into the wells of a white 96-well plate.
   • Add control antibody or sample to each well.
   • Add hFcRn-SmBiT solution to each well.
   • Cover the plate, mix gently, and incubate at room temperature.

4. Detection:

   • Prepare Lumit™ FcRn Detection Reagent.
   • Add Lumit™ FcRn Detection Reagent to each well.
   • Incubate and read the plate on a luminometer.

Opentrons Protocol:

python
DownloadCopy code
1from opentrons import protocol_api
2
3metadata = {
4    'protocolName': 'Lumit FcRn Binding Immunoassay',
5    'author': 'OpenTronsformer',
6    'description': 'Automated protocol for Lumit FcRn Binding Immunoassay using Opentrons',
7    'apiLevel': '2.11' # Adjust as per user's robot API level
8}
9
10def run(protocol: protocol_api.ProtocolContext):
11    # Labware
12    tips_300 = [protocol.load_labware('opentrons_96_tiprack_300ul', slot) for slot in ['1', '2']]
13    plate = protocol.load_labware('corning_96_wellplate_360ul_flat', '3')
14    reagent_reservoir = protocol.load_labware('usascientific_12_reservoir_22ml', '4')
15
16    # Pipettes
17    p300 = protocol.load_instrument('p300_single', 'right', tip_racks=tips_300)
18
19    # Reagents
20    tracer_lgbit = reagent_reservoir.wells_by_name()['A1']
21    hfcRn_smBit = reagent_reservoir.wells_by_name()['A2']
22    lumit_detection_reagent = reagent_reservoir.wells_by_name()['A3']
23    control_antibody = reagent_reservoir.wells_by_name()['A4']
24
25    # Distribute Tracer-LgBiT solution
26    for well in plate.wells():
27        p300.transfer(25, tracer_lgbit, well, new_tip='always')
28
29    # Distribute Control Antibody or Sample
30    # Assuming control antibody and samples are in a separate rack (update as necessary)
31    for well, sample in zip(plate.wells(), sample_rack.wells()):
32        p300.transfer(25, sample, well, mix_after=(3, 50), new_tip='always')
33
34    # Distribute hFcRn-SmBiT solution
35    for well in plate.wells():
36        p300.transfer(50, hfcRn_smBit, well, new_tip='always')
37
38    # Mix and incubate (adjust shaking and incubation as per manual)
39    protocol.delay(minutes=30)  # Incubation time
40
41    # Add Lumit™ FcRn Detection Reagent
42    for well in plate.wells():
43        p300.transfer(25, lumit_detection_reagent, well, new_tip='always')
44
45    # Incubate and read (Assuming integration with a compatible luminometer)
46    protocol.delay(minutes=5)  # Incubation time
47
48    # Luminometer reading step would be here (requires integration setup)
49
50# Validation for preventing 'NameError: name 'protocol' is not defined'
51if __name__ == '__main__':
52    from opentrons import simulate
53    protocol = simulate.get_protocol_api('2.11')  # Adjust as per user's robot API level
54    run(protocol)

Notes:

• Labware and Reagents: Ensure that the labware (e.g., tip racks, plates, reservoirs) and reagent names match what is actually used.
• Pipetting Volumes and Mixing: Adjust the volumes, mixing steps, and incubation times as per the assay requirements.
• Luminometer Integration: If your setup includes an integrated luminometer, include steps for reading the plate. This may require additional