sars-cov-2 antibody profiling

User Guide

SARS-CoV-2 Antibody Profiling: K417N, E484K, N501Y, D614G Spike S1 AntigenWI-0046V1, RELEASED AUGUST 2021

Document reference: WI-0046v12

This workflow and its products described here are for research use only and is not to be used for any other purposes, including, but not limited to, in vitro diagnostics, clinical diagnostics, or use in humans. The document and its content are proprietary to Fluidic Analytics and is intended for use only in connection with the products described herein and for no other purposes.

Fluidic Analytics products have been designed to be safe in accordance with the operating instructions. If the equipment is not used in the manner specified in the operating manual the equipment may be impaired and warranty may be void. Warranty exclusions include:

• Defects caused by improper operation or opening of the instrument

• Repair or modification done by anyone other than Fluidic Analytics or an authorized agent

• Damage caused by accident or misuse

• Damage caused by disaster

• Corrosion due to use of improper sample

The information contained in this user guide is subject to change without notice. Fluidic Analytics shall not be liable for errors contained herein or for incidental or consequential damages in connection with the performance, or use of this material.

This document contains information that is protected by copyright. All rights are reserved. No part of this document may be photocopied, reproduced, or translated to another language without prior written consent of Fluidic Analytics.

The instructions within this document must be followed rigorously by qualified and trained personnel. Failure to adhere may result in damage to the product, injury to persons or damage to other property. Fluidic Analytics shall not be liable for any damages or expenses arising directly or indirectly from the use of this product (including parts or software).

Fluidic Analytics makes no warranty of any kind, either expressed or implied. This includes merchantability for this product, and the fitness of the product for any purpose.

For Research Use Only 3

Contents

1. Intended Use 4

2. Background 4

3. Microfluidic affinity antibody profiling – assay principle 4

4. Before you start 5

5. Required equipment, reagents and consumables 7 5.1. Equipment 7 5.2. Reagents 7 5.3. Consumables 7

6. Protocol 8 6.1. SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) labeling 8 6.2. Serum measurements 10

7. Data Analysis 14 7.1. Access Fluidity Cloud 15 7.2. MAAP assay using non-linear least squares fit 16 7.3. MAAP assay using Bayesian fit 18

8. Technical support 20


1. Intended use This user guide describes microfluidic affinity antibody profiling (MAAP) against the SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) protein directly in serum samples by use of the Fluidity One-W Serum instrument.

2. Background

COVID-19 is a respiratory disease caused by the coronavirus SARS-CoV-2 that produces mild symptoms in most individuals. However, dependent on underlying health issues and other factors such as age, gender, or genetic makeup, symptoms can vary dramatically in severity. A complete and accurate immune response assessment of COVID-19 patients or vaccinated individuals based on the two key determining factors, namely antibody concentration and affinity, is key to:

• Understanding the differences in disease severity

• Assessing the strength and duration of acquired immunity

• Evaluating the mechanisms and immunological effects of virus-neutralizing antibodies

• Characterizing cross-reactivity with other, less harmful, coronaviruses

• Quantify the effect of immunization

• Evaluate different virus variants

This user guide describes affinity-based antibody profiling against the SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) for a comprehensive immune response assessment in COVID-19 patients or vaccinated individuals.

3. Microfluidicaffinity antibodyprofiling– assayprinciple

This user guide describes all steps required to characterize the immune response against the SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) directly in minimally diluted serum samples. After SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) labeling, three independent serum titration curves are measured to enable the simultaneous determination of antibody affinity and concentration.


Experimentalworkflow

03

O/N Incubation

60 min incubation at RT

Medium size range:340 min (113 min per titration curve)Large size range:570 min (190 per titration curve)

Mediumsize range:55 minLargesize range:90 min

Storage of stock aliquots at -80 °C

Antigen Labeling

01

02

Ant

igen

Lab

elin

g

Spike S1 Protein(S1) Labeling

S1 Clean-up& Labeling QC

S1 Sample Preparation

Size-range determination

Serum BackgroundMeasurements

Non-linear least squaresfit and/or Bayesian fit

KD and antibody concentration

Serum Background Measurements

Sample Measurement

Titration series 1

Titration series 2

Titration series 3Mea

sure

men

ts

Stop point

Stop point

Stop point

20 min

Dat

a A

naly

sis

04

Resu

lts


SerumpreparationandstorageFor preparation of serum samples, leave whole blood clotting for half an hour at room temperature immediately after collection. Spin down at 2,000 × g for 10 min at 4 °C.

• Immediately prepare 50 µL aliquots from the supernatant and freeze samples at -80 °C

• Only thaw samples once; additional freeze-thaw cycles will negatively impact the data

• Additives are not recommended, and addition of surfactants is strongly discouraged as it will negatively impact the data

If serum is received frozen, store immediately at -80 °C. After thawing the sample for the first time, remove the volume required for the assay, and aliquot the remaining volume into 50 µL aliquots before re-freezing at -80 °C.

When using plasma be aware that recurring freeze-thaw cycles can lead to precipitation which might negatively impact the measurements.

4. BeforeyoustartGoodlaboratorypracticewhenworkingwithserumsamples

For use of this user guide we recommend working with serum rather than plasma. Serum collected from COVID-19 patients (whether newly infected or recovered), and uninfected people who could still harbour infectious agents must be handled using a high level of precaution and at the required biosafety level for your country’s regulations. In addition, all serum samples should be handled as if capable of transmitting infectious agents.


5. Requiredequipment, reagentsandconsumables

5.1. Equipment• Fluidity One-W Serum (Fluidic Analytics

F1W0001 - SRM)

• Nanodrop (Thermo Fisher ND-ONE-W or equivalent)

• Pipettes (1000 µL, 200 µL, 10 µL)

• Zeba™ Desalting Chromatography Cartridges, 7K MWCO, 1 mL (Thermo Fisher 89934)

• pH meter (various suppliers)

• Centrifuge 5430R (Eppendorf 5428000060, or equivalent)

• Analytical Balance (Ohaus PA214C, or equivalent)

5.2. Reagents• Alexa Fluor™ 647 NHS Ester (Thermo Fisher

A20006)

• DMSO (anhydrous) (Invitrogen, D12345-10 x 3 mL)

• SARS-CoV-2 (COVID-19) S1 (K417N, E484K, N501Y, D614G) Protein: 100 µg (sufficient for affinity measurements of 100 patient samples) (SinoBiological, 40591-V08H10-100ug)

• Albumin Human Serum: 5 g (Sigma Aldrich A3782-5G)

• Sodium bicarbonate NaHCO3 (Merck S6014)

• PBS buffer at pH 7.4 (Merck P4417)

• Tween® 20 (Merck P7949)

• Glycerol (Sigma G9012)

• Ultrapure water (various suppliers)

• HCl solution at 1 M for pH adjustment (various suppliers)

5.3. Consumables• Fluidity One-W consumables – Small, 4 boxes

of chips (sufficient for 96 measurements), 1 filled cartridge (F1W0002) Alternatively: Fluidity One-W consumables – Medium, 12 boxes of chips (sufficient for 288 measurements), 3 filled cartridges (F1W0003)

• 1000 µL, 200 µL, 10 µL low-retention graduated tips (various suppliers)

• Protein Lo-Bind tubes (Eppendorf 022431081 1.5 mL; Eppendorf 0030108302 5 mL)

• Sterile filters, 0.22 µm, PVDF (sterile, various suppliers)

• 50 mL syringes, non-sterile (various suppliers)


• 5 mL Luer Lock syringe (various suppliers)

• 2 x 1 mL Luer syringe (various suppliers)

• 96-well assay plates, flat bottom clear, black polystyrene, non-binding surface (Corning® 3881)

• TopSeal™ - A PLUS, clear adhesive microplate seal (PerkinElmer 6050185)

• 50 mL Falcon tubes (various suppliers)

• Eppendorf rack (various suppliers)

• Disposable needle (various suppliers)


6. Protocol

6.1. SARS-CoV-2S1(K417N, E484K,N501Y,D614G)labeling

Requiredequipment,reagents andconsumables

Required equipment:

• Nanodrop (Thermo Fisher ND-ONE-W or equivalent)


• Zeba™ Desalting Chromatography Cartridges, 7K MWCO, 1 mL (Thermo Fisher 89934)

• pH meter (various suppliers)

Required reagents:

• Alexa Fluor™ 647 NHS Ester (Thermo Fisher A20006)

• DMSO (anhydrous) (Invitrogen, D12345-10 x 3 mL)

• SARS-CoV-2 (COVID-19) S1 (K417N, E484K, N501Y, D614G) Protein: 100 µg (sufficient for affinity measurements of 100 patient samples) (SinoBiological 40591-V08H10-100ug)

• Sodium bicarbonate NaHCO3 (Merck S6014)

• PBS buffer at pH 7.4 (Merck P4417)

• PBS-T buffer at pH 7.4 (Merck P4417, 0.05% Tween® 20 – Merck P7949); sterile filtered

• Glycerol (Sigma G9012)

• Ultrapure water (various suppliers)

• HCl solution at 1 M for pH adjustment (various suppliers)

Required consumables:




• 5 mL Luer Lock syringe (various suppliers)

• 2 x 1 mL Luer syringe (various suppliers)

• Disposable needle (various suppliers)

PreparationofrequiredsolutionsPrepare label stock solution

• Immediately before use, dissolve 1 mg of Alexa Fluor™ 647 NHS Ester in 80 µL of DMSO to prepare a 10 mM solution

• Aliquot in 5 µL portions and store at -20 °C

• Label stock solution can be used for at least 4 weeks if stored at -20 °C

• Do not re-freeze aliquots once thawed

Prepare 6-fold concentration labeling buffer

• Dissolve 8.4 g of sodium bicarbonate (NaHCO₃) in 95 mL of ultrapure water

OO Note: This might take 10 – 15 min depending on stirring speed and temperature

• Adjust pH to 8.3 with 1 M HCl if required

• Add ultrapure water to a final volume of 100 mL

• Filter using a pore size of 0.22 µm

• Store at -20 °C in 1 mL aliquots. If stored at -20 °C, 6-fold concentration labeling buffer can be used for up to 4 weeks

ProtocolSARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) labeling

• Use 100 µg of SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) at a concentration of 0.6 mg/mL (i.e., 7.8 µM) in 167 µL of sterile water

• Add 50 µL of 6-fold concentration labeling buffer and mix carefully by pipetting up and down 5 times. Do not vortex.

• Add 0.4 µL of 10 mM label stock solution to SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) and carefully pipette up and down 5 times for mixing, using a 100 µL pipette. Make up to. Do not vortex.

• Add PBS to obtain a final volume of 250 µL

• Incubate the labeling reaction at 4 °C overnight

• Protect from light


Removal of unbound Alexa Fluor™ 647 NHS Ester using a desalting column

• Connect a 5 mL Luer Lock syringe to the desalting column and equilibrate with 5 mL of PBS (pH 7.4)

OO Note: do not use labeling buffer

• Place 10 Eppendorf tubes in a rack, leave the lids open

• Connect a disposable needle to 1 mL Luer syringe

• Draw the labeling mixture into the 1 mL Luer syringe

• Remove the needle from the syringe and dispose accordingly

• Remove trapped air from the syringe before connecting it to the desalting column

• Push the sample through the column.

OO Note: It is not necessary to collect the flow-through at this stage

• Fill a second, unused 1 mL Luer syringe with 1 mL of PBS (pH 7.4) buffer

• Elute the sample by collecting 100 µL fractions in the prepared Eppendorf tubes. Read the fraction volumes using the scale of the syringe.

• The protein will elute in fractions 2 – 4, and higher number fractions will contain unbound label

LabelingQC

• Measure protein yield as well as labeling ratio

OO To determine the protein yield as well as labeling ratio, measure the absorbance of SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) at a wavelength of 280 nm and the absorbance of the conjugated label at a wavelength of 645 nm on a Nanodrop (select Alexa FluorTM 647) (ε = 96,410 M-1 cm-1 for S1)

• The labeling ratio should be close to 1 label per protein

• Typical yields are ≥15 µg

• Typical concentrations are >2 µM

OO If considerably higher label-to-protein ratios are measured, unbound label was not completely removed, and the desalting procedure should be repeated

• Check Rh of SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) on Fluidity One-W Serum

OO Dilute the protein to a concentration of 100 nM in a volume of 10 µL using 0.05% PBS-T buffer (pH 7.4)

OO Pipette 5 µL on a microfluidic chip and perform a single measurement at the 1.5 – 8 nm setting

• The typical Rh of SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) is 5.3 nm ± 0.2 nm

OO If an Rh lower than 5.1 nm is measured, unbound label was not completely removed, and the desalting procedure needs to be repeated

OO Note: The size will be different when measuring in serum

• Stock solution of the labeled protein (Alexa FluorTM 647– SARS-CoV-2 S1) is stored in PBS, containing 10% glycerol at -80 °C


6.2. Serummeasurements

Requiredequipment,reagents andconsumablesRequired equipment• Fluidity One-W Serum (Fluidic Analytics

F1W0001 - SRM)


• Centrifuge 5430R (Eppendorf 5428000060, or equivalent)

Required reagents• PBS buffer at pH 7.4 (Merck P4417)

• Tween® 20 (Merck P7949)

• Alexa Fluor™ 647-SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) probe (see section 6.1)

• Serum samples

• Albumin Human Serum: 5 g (Sigma Aldrich: A3782-5G)

Required consumables• Fluidity One-W consumables- Small, 4 boxes

of chips (sufficient for 96 measurements), 1 filled cartridge (F1W0002) Alternatively: Fluidity One-W consumables – Medium, 12 boxes of chips (sufficient for 288 measurements), 3 filled cartridges (F1W0003)






• 96-well assay plates, flat bottom clear, black polystyrene, non-binding surface (Corning® 3881)

• TopSeal™ - A PLUS, clear adhesive microplate seal (PerkinElmer 6050185)

Preparationofrequiredsolutions1% PBS-T (stock solution)• Add 0.5 mL of Tween® 20 to 49.5 mL PBS

and stir for at least 30 min at room temperature to obtain a stock solution of 1% Tween® 20 in PBS (pH 7.4) (PBS-T)

• Filter sterilize the solution using sterile filters (0.22 µm) and syringe of appropriate volume and store at 4 °C

0.05% PBS-T (working solution)• Mix 4.75 mL of filter sterilized PBS (pH 7.4)

with 0.25 mL of 1% PBS-T stock solution

• Vortex for 10 sec

• Store at room temperature

Preparation of 3.8 mM HSA in 0.05% PBS-T • Use an analytical balance to weigh 250 mg

of HSA and mix with 1 mL of PBS-T (pH 7.4)

• Mix carefully until all HSA dissolves (this might take several minutes)

• Aliquot in 100 µL portions and store at -80 °C

0.75 mM HSA in 0.05% PBS-T • Add 400 µL of 0.05% PBS-T (pH 7.4) to 100uL

of 3.8 mM HSA in PBS-T

Alexa FluorTM 647- SARS-CoV-2 Spike S1 (K417N, E484K, N501Y, D614G; probe)• Thaw one aliquot of stock solution and determine

the concentration if not known

• For the preparations of the binding curves, prepare the following three stock dilutions:

OO 1 µM, 400 nM, 100 nM and 50 nM

OO Prepare 10 µL of 1 µM antigen in 0.75 mM HSA in 0.05% PBS-T (use 3.8 mM HSA as stock solution)

OO For 0.4 µM: Take 8 µL of 1.0 µM and mix with 12 µL of 0.75 mM HSA in 0.05% PBS-T

OO For 0.1 µM: Mix 5 µL of 0.4 µM antigen with 15 µL of 0.75 mM HSA in 0.05% PBS-T

OO For 0.05 µM: Mix 6.5 µL of 0.1 µM antigen with 6.5 µL of 0.75 mM in HSA in 0.05% PBS-T

OO Keep on ice


Serum Samples• Thaw three serum aliquots to a total of 150 µL slowly on ice. Pool the aliquots and spin down 5 min at 14,000 × g.

Serum dilutions for background determination• For each serum sample, prepare 100%, 50% and 25% serum solutions using 0.75 mM HSA in 0.05% PBS-T

OO 100% serum: 0 µL of 0.75 mM HSA in 0.05% PBS-T + 16 µL of 100% serum



Instrumentset-up• Switch on the instrument (power button on the back of the instrument)

• Wait until the instrument has booted up

• Log into your account or use guest account

• Ensure the instrument is connected to the internet

• Check cartridge level. If required change cartridge as per the instrument User Guide

ProtocolPreparation of serum titration series

We recommend three titration curves with different concentrations of Alexa FluorTM 647–SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G) for the simultaneous determination of antibody affinity (KD) and concentration

• Prepare the titration series for the three affinity binding curves in a 96-well assay plate as follows:

OO Cover the plate with a sheet of adhesive microplate seal and protect from light

OO Incubate for 1 hour at 4 ˚C

1 5 nM S1

Serum 50 nM S1 0.75 mM HSA in PBS-T

Serum

A 50% 1.6 µL 6.4 µL 8.0 µL

B 30% 1.6 µL 9.6 µL 4.8 µL

C 15% 1.6 µL 12.0 µL 2.4 µL

D 7.5% 1.6 µL 13.2 µL 1.2 µL

E 3.8% 1.6 µL 13.8 µL 0.6 µL

F 1.9% 1.6 µL 14.1 µL 0.3 µL

G 0% 1.6 µL 14.4 µL N/A

H N/A N/A N/A N/A

https://www.fluidic.com/resources/fluidity-one-w-user-manual-version-380/


2 10 nM S1

Serum 100 µM S1 0.75 mM HSA in PBS-T

Serum

A 70% 1.6 µL 3.2 µL 11.2 µL

B 60% 1.6 µL 4.8 µL 9.6 µL

C 50% 1.6 µL 6.4 µL 8.0 µL

D 30% 1.6 µL 9.6 µL 4.8 µL

E 15% 1.6 µL 12.0 µL 2.4 µL

F 7.5% 1.6 µL 13.2 µL 1.2 µL

G N/A N/A N/A N/A

H N/A N/A N/A N/A

3 40 nM S1

Serum 400 nM S1 0.75 mM HSA in PBS-T

Serum

A 70% 1.6 µL 3.2 µL 11.2 µL

B 60% 1.6 µL 4.8 µL 9.6 µL

C 50% 1.6 µL 6.4 µL 8.0 µL

D 30% 1.6 µL 9.6 µL 4.8 µL

E 15% 1.6 µL 12.0 µL 2.4 µL

F 7.5% 1.6 µL 13.2 µL 1.2 µL

G N/A N/A N/A N/A

H N/A N/A N/A N/A

Size-range determination• For titration series 1, measure the 30% serum sample and the 50% serum sample [5 nM Alexa FluorTM

647-SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G; constant)] at the medium size-range setting;

OO Press “single run” OO Choose medium size-range setting (1.5 – 8.0 nm) OO Add the following labels:

• “Ligand”: Insert name• “Ligand concentration”: Serum concentration• “Protein”: Alexa FluorTM 647-SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G)• “Protein concentration”: 5 nM

OO Press “start”

• If “diffusion error (low)” occurs for any of the 2 serum concentrations mentioned above, the larger size-range setting (2.5 nm – 20 nm) needs to be selected. The serum background and the remainder of the MAAP assay samples need to be performed at the size-range setting determined in the above test.

OO Note: If the initial size-range setting was not sufficient for 30% serum and 50% serum both runs need to be repeated at the size range setting for larger samples.

OO To protect individual personal information and to comply with local and international data protection legislation, please refrain from entering any value that could be considered as personal identifiable data, be it yourself, organization or sample source.


Serum background measurements

Once the appropriate size-range setting has been selected, measure serum background:

• Purpose: each serum sample has a different level of autofluorescence. This will be subtracted from the datapoints of the titration curves later. Serum background has to be measured on the same day as the samples.

• For each measurement, pipette 5 µL of the respective serum dilutions (100%, 50%, and 25% in 0.05% PBS-T) onto a fresh Fluidity One-W chip and insert it gently into the instrument

OO Press “single run”

OO Choose the appropriate size-range setting (make sure size-range determination step was performed)

OO Add the following labels:

• “Ligand”: Insert name

• “Ligand concentration”: Serum concentration

• “Protein concentration”: 0 nM


• Measure each concentration in duplicate

Note: To protect individual personal information and to comply with local and international data protection legislation, please refrain from entering any value that could be considered as personal identifiable data, be it yourself, organization or sample source

Measurements of affinity binding curves

• Measure each datapoint of titration series 1 [5 nM Alexa FluorTM 647 – SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G; constant)] in duplicate at medium size-range setting; repeat the measurement for any potential outlier


OO Choose the appropriate size-range rate setting (make sure size-range determination step was performed)




• “Protein”: Alexa FluorTM 647 – SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G)



• Measure each datapoint of titration series 2 [10 nM Alexa FluorTM 647 – SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G; constant)] in duplicate at medium size-range setting; repeat the measurement for any potential outlier






• “Protein”: Alexa FluorTM 647 – SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G




• Measure each datapoint of titration series 3 [40 nM Alexa FluorTM 647 – SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G, constant)] in duplicate at medium size-range setting;






• “Protein”: Alexa FluorTM 647 – SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G)



Note: To protect individual personal information and to comply with local and international data protection legislation, please refrain from entering any value that could be considered as personal identifiable data, be it yourself, organization or sample source


7. Dataanalysis7.1. AccessFluidityCloudPrior to performing the analysis, please contact [email protected] for access to Fluidity Cloud.

• Accept the invitation to access the Login Portal of the Cloud Services

• Once on the Login Page, log onto the site with your username (email address) and create a password:

• When logging in for the first time, the “Experiment” page will be empty. Once experiments have been uploaded and analyzed, they will appear as a list and can be individually accessed at any time.

• To analyze new data, create a new experiment

• Select analysis method


7.2. MAAPassayusingnon-linearleastsquaresfit• Enter experiment name

• Create the experiment and then follow the on-screen steps to select the respective measurements for your experiment

• Once the measurement files are successfully selected, background subtraction will be automatically performed for each measurement, and the resulting data for each titration series will be subjected to a non-linear least squares fit to determine the following parameters:

Note: If the Fluidity One-W Serum is not connected to the Fluidity Cloud, measurement files need to be transferred manually. To do so, download measurement files from the instrument using a USB stick as per the instrument User Manual. Then use the ADD MEASUREMENT function on the Fluidity Cloud once a new experiment has been created.

Parameter Definition

KD Dissociation constant

AAntibody concentration in the sample serum

Rh, free

Hydrodynamic radius of the unbound Alexa Fluor™ 647 – SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G

Rh, complex

Hydrodynamic radius of the antibody-bound Alexa Fluor™ 647 – SARS-CoV-2 S1 (K417N, E484K, N501Y, D614G

https://www.fluidic.com/resources/?q=&type=user-manuals


• The results for each experiment will be shown as individual binding curves for each titration series

• The background-corrected values for the duplicate measurements are listed below the graph and can be viewed individually in the table below the graph

• In this table, the “status” column indicates if JSON files were included and processed correctly

• In addition, for each datapoint the number of included measurements will be indicated. As all datapoints are measured in duplicate, each datapoint should be represented by 2 included measurements. However, obvious outliers can be excluded from the analysis, and the total number of measurements that will be used for the final analysis will be adjusted accordingly.

• Using the “ADD MEASUREMENT” button, additional measurement files can be added at any point of time if required

• Analysis results as well as background corrected values for the individual measurements can be exported as a csv-file when required

Note: While outliers can be excluded from the analysis, the data will not be removed from the underlying data file to enable data tracking and auditing.


7.3. MAAPassayusingBayesianfit

• Create the experiment and then follow the on-screen steps to include the respective measurements for your experiment

• Once the measurements are successfully included, press “Run analysis”

• During the analysis, background subtraction will be automatically performed for each measurement, and the data will be subjected to a Bayesian fit. Results including the Binding curve plot, KD values as well as the binding site concentration of the unlabeled analyte will automatically be shown once the analysis is complete.

• By expanding the table, additional experimental data can be displayed. For more information please see the “Methods description” tab.


About usWe envision a world where information about proteins and their behaviour transforms our understanding of how the biological world operates, and helps all of us make better decisions about how we diagnose diseases, develop treatments and maintain our personal well-being.


• Images can be saved by clicking on the “Save Image” icon next to the image. Similarly, data can be exported by clicking on “Export Data” icon next to the image.

• Additional plots including joint and individual parameter plots, quality control data and methods description can be accessed via the individual tabs above the table. For more information on these please refer to the “Methods description” tab.

• The background-corrected values for the individual measurements are listed below the graph

• In this table, the “status” column indicates if measurements were uploaded and processed correctly

• In addition, for each datapoint the number of included measurements will be indicated. Obvious outliers can be excluded from the analysis, and the total number of included measurements that will be used for the final analysis will be adjusted accordingly.

Note: While outliers can be excluded from the analysis, the data will not be removed from the underlying data file to enable data tracking and auditing.

• Using the “ADD EXPERIMENT” button, additional measurements can be added at any point of time if required

8. TechnicalSupportFor technical or data analysis support, please contact Fluidic Analytics Technical Support at [email protected].

Copyright © 2021, Fluidic Analytics. All rights reserved. Fluidic Analytics and Fluidity are trademarks of Fluidic Analytics. Other product names, service names, logos and/or company names used are trademarked and copyrighted properties of their respective owners and/or licensors. For Research Use Only.

[email protected]

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sars-cov-2 antibody profiling

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