We offer diverse custom PolyHRP conjugations with your own antibody, antigen, etc. If
substance you wish to have labeled with PolyHRP has available at least one free primary
aminogroup (provided not hidden, i.e. in free access onto the surface of your given
molecule), we can try to couple it.
PolyHRP is a perfect enhanced label candidate for coupling with detector anti-analytes in
two-site assays. These are both detector antibody in double antibody sandwich for an
antigen and detector (second) antigen in double antigen sandwich for specific antibody - the
most sensitive and specific antibody assay scheme employing immobilized and labelled
antigen. Custom PolyHRP conjugations are priced depending on starting amount* of
material for labeling as follows
Custom PolyHRP Conjugations pricing
PolyHRP conjugation, in contrast to e.g. biotinylation, is very scale-sensitive process.
Unusual stoichiometry of PolyHRP conjugate synthesis makes smaller volume conjugations
technically very difficult.
That is why the smallest starting amount of protein (antibody, antigen, etc.) for trial custom
PolyHRP conjugation is 10-20-30 mgs. On the other hand this makes a visible economical
advantage: in positive case customer obtains at least 20 to 30 mgs (and probably larger
quantity) of e.g. enhanced high performance Antibody-PolyHRP conjugate which is a good
production amount sufficient for making many thousands of ELISA test kits (roughly
estimated 10.000 to 40.000 standard 96-well micro-titer plates depending on conjugate
working strength dilution).
10 mg at 5-10 mg/ml is an optimal minimum starting amount for the effective PolyHRP labeling - in general.
We have been trying to decrease this amount as it is often a restraint to many research and small
industrial and semi-industrial customers and can currently go down to 4-5 mg IgG/F(ab')2
and even to 2-3 mg with smaller 20-25-30 kDa peptide/glycoprotein antigens.
Yield will depend on (i) specific characteristics and behaviour of your given substance in
course of the synthesis that are not always smoothly predictable and (ii) desired grade of
HRP homopolymer in signal-generating conjugate moiety. With good antibody (whole IgG)
we can ensure nominal yield at a level of at least 1 mg GPC/SEC purified Antibody-PolyHRP40
conjugate per 1 mg starting material. In many cases, with good antibodies, we
can provide up to 40-50% better yields. Yields achievable in PolyHRP80 conjugations are
significantly lower compared to PolyHRP40. lately we have de facto ceased conjugations of
the large molecules like IgG with PolyHRP80. We also discourage customers from PolyHRP20
conjugations as PolyHRP20 will unlikely perform better than PolyHRP40 and desired PolyHRP
grade has no influence on the overall price. We basically recommend starting (and in most
cases ending) with PolyHRP40 that generally gives both reasonable yield and good performance.
Antibody conjugations. Detection activity / performance is essentially better with
antigen-binding F(ab')2 than with whole IgG antibody. Material
closer to chromatographically pure 150 kDa IgG antibody is clearly better material for
coupling compared to preparation containing heavier non-covalent IgG polymers or other
molecular aggregates and complexes.
Optimal concentration of protein (antibody as IgG or F(ab')2, antigen, peptide, etc.) for labeling is 5-
10 mg/ml. This also means that material to be labeled should be stable at these
concentrations in solution. If material supplied for labeling in smaller concentration, we will
concentrate it. If it tends to precipitate or auto-polymerize at higher concentrations,
a part of the material that we will lose through concentrating will not be recovered and respectively
incorporated in conjugate
anyway. Also, avoid presence of sodium azide in your material, when possible. Any extra
processing of your material aimed at making it suitable for conjugation may cause
additional material losses.
We strongly recommend giving us available information on general physico-chemical
characteristics of your substance as size/M.Wt., i.p., other data that can be helpful in planning
and running custom synthesis.
We do not recommend ordering custom PolyHRP conjugation, if you are not sure that your
material works well as anti-analyte moiety in conventional HRP conjugate, i.e. if you have
no available experimental data evidencing satisfactory performance of your HRP conjugated
material in existent ELISA or related assay test system.
We will run custom synthesis with your material essentially blindly - having no
control over its functional performance, and will deliver prepared custom conjugate with no
functional QC release.
We will charge extra €500,00 for making extra functional (ELISA test) quality control with
synthesized material. In this case we ask that customer provides us with all necessary for using
in specific QC release procedure materials, e.g. capture Antibody, Antigen
Standard/Calibrator and (optionally, however strongly recommended)
standard/conventional detector Antibody-HRP conjugate of comparison that is currently
being utilized (exampled for the case when custom Antibody-PolyHRP conjugate shall be used in double antibody
sandwich (ELISA) test system).
Turnaround time is from 1 (rather seldom) - 2 to 4-6 weeks and will depend on the current
production schedules. We recommend choosing in coordination with us a time window that
would optimally fit to running conjugation for you. This may suggest coordinated preliminary
planning in approx. 1-2-month/week period - provided that you do not need custom conjugate badly.
Custom PolyHRP Conjugation orders may be a subject to delayed/collected processing as
we usually try to plan, depending on current production schedules, more than one specific
custom conjugation per regularly scheduled larger synthesis run.