Lateral Flow

Development and
Manufacturing Journey

Shaun Phillips
R&D Senior Scientist

Quality Partnership Results Innovation

Overview

 Contract R&D overview

 Development phases

 Technical aspects to be considered during

development

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Overview
Reliable contract R&D developers need to be able to offer a service that
address needs in the following key areas:
Experience
 Multiple detectors labels
 Varied test formats and platforms
 Worked with a variety of sample matrices

Cost effectiveness
 Experience with variety of test strip materials
 Established relationship with materials suppliers
 Access to existing equipment for development manufacture
 Efficient manufacturing process

Manufacturing transfer
 Manufacturing capabilities at scale required

Quality & Regulatory

 A comprehensive and strategic paperwork trail is vital
 Design control to ISO 13485 and
other applicable standards
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Product Development Process for IVD’s
Key deliverables for each phase

Optimisation & Design

Prep for Post Market
Feasibility Characterisation Verification &
Launch Activities
Validation

Assay
optimisation,
detailed product Final
specification, manufacturing
QC process documentation,
Project team,
development verification and
Business Review

detailed Regulatory
Design Review

Design Review

Design Review
Design Review
preliminary validation
project plan, documentation
stability studies, activities,
draft submitted to
risk final V&V Implementation
specification, appropriate
management report, final QC of post market
regulatory authority
design freeze, specification, vigilance and
pathway,
manufacturing final BOM, surveillance
feasibility Regulatory
documentation, DMR, final
report, approval
validation packaging and
updated obtained
master plan, labelling,
business case
regulatory plan, marketing plan,
packaging updated
development, business plan
assay
characterisation
Design reviews
Review all
aspects of
project
 Hold cross functional
design reviews at the end
of each development
phase

 Internal design review & Minutes

external design review Design control Assess risk
recorded
(project team and
customer)

Multi
functional
team

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Phase I – Feasibility
Key Deliverables

Identify project team

Set up design history file

Detailed project plan including detailed development

plan, V&V plan

Manufacturing plan, initial supplier assessment, risk

management plan and resource plan

Draft product specification and feasibility report

Identify regulatory pathway

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Design history files
Experimental
work &
technical
 Established for every reports

project

 Records all documentation

generated for the duration
of the project including Minutes of
SOPs,
product specifications Design design reviews
validation
history file &
documentation
(design inputs/outputs) inputs/outputs

 All technical data and

reports for each phase

Project Gantt
charts

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Technical considerations in feasibility phase

Assay
format

Prototype Antibody
evaluation selection

Feasibility

Test strip Assay

materials label

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Material selection in feasibility
Antibody selection
 Commercial antibody or in-house
 Monoclonal vs polyclonal or combination of both
 Methods of purification
 Antibody pairing
 Lot to lot variability

Label of choice
 Gold / latex / paramagnetic / fluorescent
 Conjugation conditions
 Conjugate stability

Material selection
 Nitrocellulose membrane choice and flow rate
 Conjugate pad
 Upper wick
 Housed or un-housed

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Importance of nitrocellulose quality
High quality
 More even surface for protein binding
 Less aggregates
 Good stability
 Consistent batch-to-batch
 Optimum surface area for even
loading
 Better functionality

Poor quality
 Variable surface area, uneven loading of protein
 Aggregates
 Poor stability
 False positives
 Batch to batch variation
 Varying signal intensity

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Critical test strip materials to evaluate in
feasibility
Conjugate pad

 Typically use glass fibre materials

 Selected material needs to:
 Absorb detector reagent consistently
 Release detector consistently
 Maintain stability of conjugate for shelf-life of test
 Low protein binding
 Good tensile strength

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Critical test strip materials to evaluate in
feasibility
Sample pad

 Typically use cellulose papers or glass fibres

 Selected Material dependent on Sample Matrix
 May be required for sample separation/filtration
 Ensure all sample volume absorbed
 Chemical treatment
 Good tensile strength
 Low cost

Upper wick

 Typically use cellulose papers

 Ensures flow in assay so absorbency critical
 Low cost

Backing card

 Structure for strip

 Adhesive quality very important
 Accurate dimensions

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Critical materials – assay housings

Many varieties – internal structure is critical to assay performance – this is not

appreciated by many customers

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 Testing methods – gold conjugate checkerboard
BBI employ a number of testing methods to make the design control process more efficient

Nitrocellulose
CAMAG
Ab 1 Ab 2 Ab 3 Ab 4 Ab 5 Ab 6 Ab 7 Ab 8 Ab 9 Ab 10 Ab 11 Ab 12
values
Neg Pos Neg Pos Neg Pos Neg Pos Neg Pos Neg Pos Neg Pos Neg Pos Neg Pos Neg Pos Neg Pos Neg Pos
Ab 1 342 683 22.4 19.4 26.2 16.6 51.3 59.1 38.3 60.3 45.9 42.3 64.7 117 45.2 413 65.5 178 388 663 89.9 134 9.9 32.5
Ab 2 341 302 35.4 21.1 15.9 40.4 47.3 69.4 37 30.6 18 34.7 85.3 30 51.4 53.1 29.7 58.8 284 232 78.4 63.5 19.5 7.4
Ab 3 103 410 13.9 20.2 39.5 17.3 33.2 42.9 54.4 41.4 31.8 39.2 53.5 61.6 27.4 62.2 24.4 35.8 320 480 68.6 80.8 16.8 14.8
Ab 4 205 391 22.8 27.6 64.7 24.8 31.7 63.3 58.2 50 61.3 38.7 115 89 44.2 63.4 31.7 42.2 252 441 110 79 15.5 41
Ab 5 472 253 30.9 25.4 25.6 35 32.6 59.4 25.5 23 29.1 58.8 12.5 18 33.3 21.4 34.2 52 320 457 28.2 50.5 15.3 80
Ab 6 382 331 19.8 12.2 35.4 14.3 45 45.6 71.9 62.9 53.5 37.1 46.1 29.1 43.5 32.3 22 32.2 342 195 93.7 108 17.4 169
Ab 7 268 361 28.2 42.2 41.5 57.6 53.2 116 48.3 36.1 94.8 106 64.6 108 64 170 66.6 103 267 455 55.7 271 38.4 157
Ab 8 283 525 15.4 13.2 18.1 49.9 34.5 43.4 47.9 56.1 28.9 18.8 74.2 89.1 57.3 343 42.4 57 144 236 90 30.4 11.6 141
Gold Conjugate

Ab 9 354 536 126 102 18.8 22.3 35.9 45.8 55.4 39.7 90.6 73.2 70.1 66.4 93.3 63.8 56 75.3 182 313 82.7 88.6 16.8 43.9
Ab 10 379 609 40.6 26.7 23.6 11.3 23.6 37.7 23.8 21.5 48.8 35.4 84.5 107 55.5 353 36.4 147 359 428 41.5 70.9 11.1 38.2
Ab 11 550 505 30 39.3 49.8 31.5 32.5 49.9 13.8 20 145 171 63.1 191 230 131 116 93.8 245 129 477 479 38.9 54.9
Ab 12 286 60 32.6 15.4 11.4 11.5 20.1 47.4 18 10.7 28.7 15.1 58.5 94.5 15.9 35.9 22.4 47.7 566 169 32.8 57.7 8 110
Ab 13 68.6 653 14.3 11.2 7.3 12.9 47.5 74.5 70.2 120 92 61 106 215 88.7 308 13.7 33.4 352 659 25.9 40.6 5.6 51.1
Ab 14 708 665 101 199 62 84.9 85.5 94.6 185 207 279 394 456 135 331 303 210 148 673 700 291 206 13.6 181
Ab 15 256 354 23.8 11.2 47 25.2 37.6 48.5 42.5 44 37.7 46.5 59.9 80.1 40 47.5 32.1 28.1 489 468 59.5 82.2 15.2 70.4
Ab 16 486 344 13.5 8.4 38.2 20.3 30.4 31.8 58.3 48.3 49.1 38.9 55.8 81.5 49.8 23.4 32.3 14.1 345 471 51.2 76 10.5 5.1
Ab 17 403 226 9.2 51.1 46.6 18.6 57 50.3 57.7 29.8 42.1 59 106 117 110 49.9 59.9 26.2 537 524 25.6 36.6 6 10.6
Ab 18 578 472 14.2 8.6 5 12.2 39.2 37.4 47.1 243 58 44.5 49.9 101 51.3 82.4 55.5 19.9 460 242 51.1 9.8 4.3 7.8
Ab 19 277 249 15.7 10.5 6.3 8.2 36.2 47.2 48.9 73.4 52.6 77.7 45.9 71.7 61.4 36.7 73.5 38.6 521 313 41.6 36.6 7.8 36.9
Ab 20 127 308 14.3 6.4 9.9 16.3 30.8 42.6 50.2 39.8 40.9 55.6 28.7 55.6 20.3 49.1 42.3 24.5 73 312 50.2 70.5 12 34.5

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Testing methods – gold conjugate checkerboard
(simplified)
Nitrocellulose
CAMAG values
Ab 1 Ab 2 Ab 3 Ab 4 Ab 5 Ab 6 Ab 7 Ab 8 Ab 9 Ab 10 Ab 11 Ab 12

Ab 1 Poor Poor Poor Poor Poor Poor Poor Good OK Poor Poor Poor
Ab 2 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 3 Good Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 4 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 5 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 6 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor OK
Ab 7 Poor Poor Poor Poor Poor Poor Poor OK Poor Poor Good OK
Ab 8 Poor Poor Poor Poor Poor Poor Poor Good Poor Poor Poor OK
Gold Conjugate

Ab 9 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 10 Poor Poor Poor Poor Poor Poor Poor Good OK Poor Poor Poor
Ab 11 Poor Poor Poor Poor Poor Poor OK Poor Poor Poor Poor Poor
Ab 12 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor OK
Ab 13 Good Poor Poor Poor Poor Poor OK Good Poor Poor Poor Poor
Ab 14 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor OK
Ab 15 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 16 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 17 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 18 Poor Poor Poor Poor OK Poor Poor Poor Poor Poor Poor Poor
Ab 19 Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor Poor
Ab 20 Poor Poor Poor Poor Poor Poor Poor Poor Poor Good Poor Poor

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Assay kinetics
It is important to monitor throughout development even for a visual assay

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Assay performance
 Hundreds of factors interact in one step assay
 All need to be balanced to ensure maximum assay performance

Assay
Specificity

Assay
Sensitivity

Once critical materials are identified – they require optimisation

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Phase II – Optimisation and Characterisation
Optimise all assay components to ensure robust design

Sample pad
manufacturer,
sample matrix,
treatment
Assay
characterisation Upper wick
to establish manufacturer,
performance of absorbency,
size
the assay prior to
V&V

Batch to batch
variability,
security
supply, Conjugate pad
Strip volumes manufacturer,
dimensions required material,
width, length, treatment, size,
overlaps, tensile
tolerances strength, drying
conditions

Coated
Housings
nitrocellulose
pinch points,
line positions,
sample well
coating
volume,
conditions,
plastics,
drying
design
conditions
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Example of buffer optimisation – reduced
factorial
Main Effects Plot (data means) for BO Test Mean
C omponent lots pH BS A %

340

330
Mean of BO Test Mean

320

A B 8.0 8.4 1.08 1.32

S ucrose % Heparin % TRIZM A mM

340

330

320

35.9 43.9 3.6 4.4 360 440

Tw een20 %

340

330

320

0.072 0.088

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Phase II – Optimisation considerations
key deliverables to be completed:

Assay Optimisation, technical work and final report

QC panel development, evaluation, stability and draft documented QC process

Equipment, facilities and utilities qualification

Generation of pilot batches (including material diversity)

Manufacturing specification

Informal stability

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Phase II – Optimisation considerations contd.

Characterisation of assay

Risk management

Packaging development and supplier assessments finalised

Finalise manufacturing documentation and design freeze

Validation master plan (Process, Performance evaluation, stability etc.)

Packaging design finalised

Design review 2

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Phase III – Design Verification and Validation

Final QC Formal stability and Updated BOMs

Specifications transport trial

Verification and
validation report-
DMR, DHR and Finalised packaging
including
method validation and labelling
performance
evaluation

Three validation
Process verification Final risk
batches to
and validation report management report
manufacturing scale

Design review 3

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Phase IV – Preparation for Launch
Finalised design dossiers,
technical files and other regulatory
documentation

Regulatory documents submitted

to appropriate authority

Marketing plan, sales and customer

support plan in place

Regulatory approval obtained

Close out of development

Design Review 4

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Phase V – Routine manufacture / post market
surveillance

Implementation of
post market
surveillance

Registration of
Customer
product in new
feedback
territories

Completed phase Risk management

checklists review

Customer
feedback

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Summary

 Design control phases

 Key technical activities required in each
phase
 Highlighted critical test strip raw materials
 Testing methods used at BBI to determine the
best combinations of raw materials and
antibodies - ensuring maximum performance
and robust design

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Raw Materials Contract Development & Manufacture
 Antibodies  Lateral Flow Test Development
 Antigens  Lateral Flow Test Manufacture
 Enzymes  Full Regulatory & Post Market Support
 Gold Nanoparticles  Sensitivity Enhancement Technology
 Serum & Plasma  Platforms
 Cell Culture Reagents  Readers

www.bbisolutions.com info@bbisolutions.com

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