고순도
대량 정제 Refolding 기술 외에도,
알츠하이머와
HIV(면역결핍증) 및 각종암에 대한, 질병의
신약 개발에 중요 요소인 Enzyme을 공급한다.
The universal refolding procedure
The core technology of ProteomTech is the
HT-REP(High-Throughput Refolding and Purification) technology
developed by Dr. Xinli Lin. Dr. Lin has been working for
nearly 15 years developing this technology. It is a
universal procedure for refolding or re-naturing of
denatured proteins, especially proteins expressed in E.
coli as inclusion bodies. Because of the long-term
development, the HT-REP procedure is mature enough for large
scale refolding, simple enough to tackle the entire human
genome, robust enough for industrial scale operations, and
economical enough for commercial applications.
MethodThis unique procedure consists of six
steps:
- Growing E. coli carrying an expressed plasma,
- Inducing the synthesis of the expressed protein as
inclusion bodies,
- Purifying the inclusion bodies with freeze-thaw and
detergent washing,
- Solublizing the inclusion bodies in a 8 M urea buffer,
- Refolding the denatured protein to its native form, and
- Purifying the refolded protein using column
chromatographic procedures.
Advantage of this method of high-throughput
statement
- Growing E. coli is faster and less expensive
compared to mammalian, insect, and yeast cultures.
- Some proteins are harmful to the host when expressed in
their native forms, thus insoluble expression is the
preferred way to obtain large quantities of the recombinant
proteins. In fact, most important proteins are highly
regulated in mammalian cells, partly because over-expression
of these proteins in their native form is harmful to their
host. Thus expression as inclusion bodies is the best way to
purify large quantities of some commercially important
proteins, such as the beta-secretase involved in Alzheimer’s
disease and HIV protease in AIDS.
- High levels of expression can be achieved for most
proteins. 200 to 400 mg of inclusion bodies per liter of
bacterial culture is routinely obtained when cells are grown
in shakers.
- The inclusion bodies are easily purified to greater than
90% purity with a simple freeze/thaw and detergent washing
procedure, making parallel production of many proteins
possible.
- The HT-REP technology developed in this lab is simple
enough that the high-throughput purification of recombinant
proteins can easily be achieved.
Uniqueness of the methodDr. Xinli Lin is the sole
inventor of the HT-REP technology, patented by the Oklahoma
Medical Research Foundation (patent pending), and the patent
has been licensed to ProteomTech. No other similar procedure
has been published or patented.
Following is a
partial list of human proteins made using the proprietary
refolding method.
- Pepsinogen
- Pepsinogen
- Rhizopus-PP
- Thermopsin
- HIV protease
- SAP
- C-II
- SK
- Plasminogen
- Cadosin A
- BACE1
- BACE2
- SAP1
- SAP2
- SAP3
- SAP6
- Fez1
- Rab5
- POMC
- TCCR
- CDC42
- ZNF230
For more information on the
above proteins, download our document click
here
For industrial and
academic labs that express proteins as inclusion bodies in
E. coli but have a difficult time refolding and
purifying the proteins, ProteomTech is the source for solving
your problems. For proteins expressed in E. coli in
soluble forms but suffer from low yield and difficult to
purify in large scale, we can help solve the problem by
expressing the proteins in the inclusion body form, and using
our "Universal" method to refold and purify the proteins.
Academic discount is available and the terms and fees of the
service contract are subjected to negotiation. Please contact
us for more information.
Polyclonal and monoclonal
antibodies are produced for each purified, native protein.
Polyclonal antibodies (affinity purified) against E.
coli expressed, purified BACE1 and BACE2 are currently
available.
Recombinant proteins purified
from E. coli are the best sources for structural
determination using either X-ray crystallography or NMR. Our
goal is to purify 500 to 1000 human proteins using our
high-throughput refolding method in the first year of
operation. These proteins will be contracted out to a
structural genomics company or academic labs for structural
determination. Please contact us for details in collaboration
or contract.
With the
high-throughput refolding and purification, the company will
build a protein bank containing thousands of proteins within
a few years. The protein bank, and the antibody bank derived
from it, will be used to manufacture microarray chips for
commercialization. The microarray chips will be manufactured
either in house or contracted out to a protein chip company.
Interested parties may contact us.
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