Aybar Ecotechnologies, a company dedicated to the development of new
technologies, has developed a new technique (the AP Aybar Protocol) for the
purification of macromolecules with ionic functional groups using the High
Performance Liquid Chromatography (HPLC). This technique has been
successfully tested in several enzymes; it could be readily applied to any kind of
polypeptides (like interferon) and protein complexes like immunoglobulin
(vaccines).
Older methods, like Hidroxylapatite Chromatography (HA) and other molecular
sieving techniques, have numerous drawbacks; Which include i) high
consumption of buffer, ii) low flow rate making the process time consuming, iii)
loss of the protein activity being purified during the purification process, iv) high
dilution of the protein extract after this chromatographic step, v) difficult or non
regeneration of this step’s chromatographic column, vi) low purification fold, thus
producing a protein extract with a high content of debris polypeptides and
proteins, and vii) due to factor vi, short life span of the affinity chromatography
column for the final step in the purification procedure. All these drawbacks
amount to an extremely high cost of the purification step in the production of
enzymes, interferon, antibodies, vaccines, and many other polypeptide peptide
biomolecules.
Aybar Ecotechnologies wishes to propose, to a Company interested, an
alliance to use the Aybar Protocol (AP) to purify proteins and related
polypeptides. The AP is a new procedure for the purification of macromolecules
with ionic functional groups, mainly proteins, using High Performance Liquid
Chromatography (HPLC). This new technique makes the process of protein
purification faster, simpler, less expensive and gives a higher yield.
The cost of purification of proteins is high, especially for the therapeutic proteins,
ranging from 50% to 80% of the overall production cost. The cost reduction
derived from the use of this technique in the purification of proteins is significant.
This relative lowering of cost to produce proteins will give the user of the AP
procedure a great advantage in the market.
With the help of a table let us make a comparison between a method commonly
used in the first stages of protein purification, Hydroxylapatite-Matrix
Chromatography (HA) and the Aybar Protocol (AP) in the purification of the
enzyme HGPRT’ase.
FEATURES AP HA
Yield (Total activity recuperated after
purification during this step) 130% 7%
Degree of Purification During this step 23 Fold 5 Fold
Stages 1 Step 5 Steps
Time required to complete the Process
(for the processing of 60 pounds of
yeast)
4-6 Days 6 Months
Dilution of protein in the effluent 3-5 Fold 50 Fold
Regeneration of the AP column 3 hours / 250ml Matrix
Several
days /
250ml
Matrix
Lifetime of the affinity Column (GMPSepharose)
Multiple times without
losing it’s binding
capacity
3 Times
Volume of elution buffer 1-3 Times (column’s
void volume)
30 Times
(column’s
void volume)
We are interested in finding a company to form a partnership with us in the use
or exploitation of this technique. As a way to show our confidence in the
performance of the AP technique we propose an arrangement in which you
would only grant us a percentage of the savings in purification costs that the use
of the technique will give you. But we are open to any kind of agreement you may
suggest for using our technique
Let us emphasize some aspects of our innovation.
- To use our protocol it is not necessary to invest additional money in
machinery, facility, or any other investment. You only need a Fast Protein
Liquid Chromatography instrument (From Pharmacia), or any system with the
same characteristics for automatic programmable injection, programmable
buffer gradient, pumping system, and preparatory chromatographic columns
packed with suitable ion exchange chromatographic resin. - AP technique can increase the yield of protein purification (activity
recuperated after purification) several times. - AP procedure consumes less buffer solution than any other purification
chromatographic procedure. - AP technique produces such a high degree of purification in a single step that
the next step in the purification procedure, the affinity chromatography step, is
improved by prolonging the life time of the affinity column. - The overall purification time involved in the AP procedure is considerable
shorter than in other procedures. - In the AP technique, the protein purification process can be automated in a
semi-continuous fashion. - All these features make the AP procedure a technique that dramatically
reduces the purification cost of macromolecules of protein nature.
As to provide you with a faithful evidence of the positive characteristics of AP, we
would be glad to implement the procedure in one of your protein products, this
we could perform, if you provide the starting material and your facilities, at no
cost to you.
We look forward to hearing from you in the near future.
Sincerely yours,
Diógenes Aybar, Ph.D.
President
Aybar Ecotechnologies