| 1 |
How do you purify the antibodies? |
| |
In most cases we follow a protocol based on Protein A or G affinity chromatography. |
| 2 |
How do you determine the quantity of the
purified antibody? |
| |
We determine protein concentration by either Lowry
Protein Assay against a BSA standard or
OD-readings. |
| 3 |
How do you check purity and quality of the
purified antibody? |
| |
We check purity and integrity of the purified antibody
routinely by a denaturing and reducing SDS-PAGE. Furthermore,
we can offer a range of optional quality and activity
tests such as flow cytometry or ELISA. Any other test
system can be established according to the customer's
needs. |
| 4 |
How do you determine the production rate
of a given hybridoma? |
| |
We usually determine productivity by an evaluation project
(one liter). Depending on the outcome we recommend different
protocols in order to purify the desired amount of antibody
in the most appropriate way! |
| 5 |
What types of antibodies
do you purify? |
| |
We can purify antibodies from sera, cell culture supernatants
or ascites. In most cases, we purify antibodies of the
IgG-subclasses, however, protocols for
IgA and IgM are also available. |
| 6 |
Do you deliver the antibody in a buffer containing
azide? |
| |
No, as azide might interfere with some special down-stream applications based on living cells. However, we can dissolve the antibody in PBS containing 0,02% sodium azide, if required. |
| 7 |
How long do you need to purify my antibody? |
| |
Since we deal with living cells timelines of a project
basically depend very much on the proliferation rate
of the hybridoma or cell line in question. As a benchmark
for a well proliferating cell line you can refer to
the following timelines:
Sera, Ascites and purification starting with supernatant
provided by the customer:
1 - 2 weeks depending on plant utilization.
Evaluation projects and cell culture based production
and purification based on supernatant volume of up to
2 liter:
4-6 weeks
Larger volumes :
Please inquire
|
| 8 |
Can you purify my antibody from serum-free
media? |
| |
We usually cultivate the hybridomas in serum-free medium. |
| 9 |
How do you avoid co-purification of bovine
antibodies originating from fetal calf sera (FCS)? |
| |
Since standard FCS does contain bovine antibodies in
varying amounts and Protein G does bind bovine IgG, our
cell culture schemes avoid the use of FCS! |
| 10 |
Can you produce antibodies from ascites? |
| |
Yes, we can. Please be aware of the fact, that for very
fatty and viscous ascites the purification might involve
additional clearing and pre-purification steps which might
prolong timelines of ascites purification projects. |
| 11 |
We have a transgenic CHO-line producing single-chain
variable fragments (scFv)? Are you able to purify it? |
| |
This depends on the exact features of your project!
If an affinity tag is included into the scFv-framework,
we can purify it based on this affinity tag. In case you
do not have such a tag, it would depend on the availability
of your antigen. Please inquire
for more details. |
| 12 |
Can you label antibodies? |
| |
Yes, we offer labeling of antibodies based on all current
dyes. Since we can offer a quality control for biotinylation
(determination of average number of biotin per antibody
molecule) most of our customers actually choose biotinylation.
FITC and PE are the second and third most popular labels,
with FITC being a good choice for intracellular staining
due to its small size. However, its relative fluorescence
makes it rather dim and therefore PE is a good choice
for all applications with the need for bright signals. |
| 13 |
Can you produce Fab or F(ab')2 fragments? |
| |
Yes, we offer fragmentation of antibodies. Since efficiency
of fragmentation very much depends on the isotype of your
antibody, please inquire
for more details . |
| 14 |
Why do you offer a Mycoplasma-screening at
the initial stage of the project? |
| |
Mycoplasma are the major obstacle for cell culture!
Furthermore, they often remain undetected. Since we do
not want to risk any sort of reduced proliferation and
production rate due to a possible contamination we initially
screen all incoming cells by PCR. Thereby we ensure fast
and productive processing of your order! |
| 15 |
I am very much concerned about confidentiality
since my antibody represents important IP to me. How do
you ensure this confidentiality? |
| |
We of course understand this very important issue of
our customers. We have established a scheme which guarantees
you absolute privacy: Although we freeze cells at an initial
stage of the project as a back-up system, we can provide
you with a certificate of destruction of these vials as
well of the flow-through after purification and finalizing
of the project. Furthermore, we are happy to sign an MTA
or a CDA if necessary. |
