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Antigen-Down
Microtiter Plate Immunoassay Sample Quote
Introduction
In an antigen-down immunoassay,
the analyte is coated onto a 96-well microtiter plate
(rather than an antibody) and used to bind antibodies
found in a sample. When the sample is added (such
as human serum), the antigen on the plate is bound by
antibodies (human IgE for example) from the sample,
which are then retained in the well. A
species-specific antibody (such as goat IgG anti-human
IgE) labeled with HRP is added next, which binds to the
antibody bound to the antigen on the plate. The
higher the signal, the more antibodies there are in the
sample. Antigen-down assays can be configured as
rapid tests and are often used to diagnose allergy
conditions - routinely, a patient's blood is tested
against different allergens to see if the person has
antibodies to that allergen.
If
the target analyte and necessary antibodies are
available, assay development generally costs between
$30,000 - $100,000 and usually takes 4-8 months.
Proposals are often broken down into 4 phases, which
detail time and cost estimates. Payments are often
structured with a front fee of 30% of the estimated
total, and milestone payments disbursed thereafter,
which can be paid after each phase or monthly.
When
the assay has been developed, we normally supply
customers with reagents for 25 immunoassay kits. They
can be supplied as individually packaged kits or in bulk
packaging (for a detailed description of the reagents in
an ELISA kit, see What
Are the Typical Components in an ELISA?). When
stored at 2o-8o C, each kit may
have a shelf life of at least 6 months. If you
need more kits, ICT will manufacture the components as
you order them (this usually requires a 3-6 week
lead-time). Once developed, the cost for additional kits
typically ranges from $50 to $300 each (with a minimum
manufacturing charge of $2,000). This price will
be determined during development and depends on the
expense of the kit components, the type of packaging
required, and the volume ordered. If you will be
testing samples at different locations, we can ship the
assay kits directly to the other laboratories (also
see Kit
Manufacturing).
This
sample proposal is meant to answer basic questions, and
to act as a project guideline. Depending on your
specific project, the amount of antigen required may
vary, conjugations may be necessary, the antigen may
need to be purified, sample preparation steps may be
necessary, and the length of time for the project will
vary. In order to answer some of your
questions, or to prepare a specific quote for your
project, please review the Assay
Development Questionnaire, and then contact us.
All prices are in US dollars.
SAMPLE
PROPOSAL
Antigen-Down Microtiter Plate Assay Development
Required materials and
reagents to begin assay development:
- Analyte.
- Antibodies.
- Analyte negative
samples.
- Analyte positive
samples.
Phase I: Reagent
Preparation and Initial Assay Titration
.............$
Time: 4-8 weeks
- Labeling of specific
polyclonal antibody (such as goat IgG anti-human IgE)
with HRP) for assay signal generation.
- Determination of a
working titration of capture antigen and detection
polyclonal antibody conjugate.
- Development and
optimization of ELISA plate coating procedures for
optimal antibody capture.
Phase II: Assay
Optimization
.
...........
...........$
Time: 4-14 weeks
- Selection of proper
diluents for conjugate signal generation.
- Development of special
additives to eliminate sample matrix effects such as
interference and non-specific binding.
- Construction of a
standard curve to mimic performance characteristics
of the sample matrix.
- Development of a
functional assay protocol within the target
sensitivity range of the target antibody.
- Elimination or
modification of extraction or sample preparation
steps.
Phase III: Assay
Validation
..............
....
.......$
Time: 4-5 weeks
- Definition and
documentation of assay performance characteristics
essential for optimal assay utility (such as
sensitivity and precision).
- Documentation of sample
performance parameters such as dilution and linearity
and nonspecific background signal generation.
- Fine-tuning of specific
assay components and incubation protocols to meet
final performance requirements.
Phase IV: Production of
25 Finished Kits
.......................
...$
Time: 3-4 weeks
- Quality control
assessment of final components.
- Final assembly,
packaging, and delivery of components for 25 ELISA
kits.
Total Assay Development
Cost
... range of $30,000 - $100,000
Estimated Time: 15-31 weeks |
