Applications of Immobilized enzymes in Health Care

Presented by: Gautam Pradeep Madhab kamala

13BT60R17 1/11/2013

Jack Andraka
Jack Andraka is a Maryland high school sophomore who at age 15 invented an inexpensive and sensitive dipstick-like sensor for the rapid and early detection of pancreatic, ovarian and lung cancers. After a close family friend died of pancreatic cancer, Jack (then a ninth grader) became interested in finding a better early-detection diagnostic test.

The human mesothelin-specific antibodies were mixed with single-walled carbon nanotubes .

It is used to coat strips of ordinary filter paper. This made the paper conductive.

The optimal layering was determined using a scanning electron microscope.

Any change in the electrical potential of the sensor strip (due to the changing conductivity of the nanotubes) was measured, before and after each application. Cell media spiked with varying amounts of mesothelin were then tested against the paper biosensor.

Can carbon nanotubes be used to immobilize biological molecules or enzymes?

TYPES OF ENZYME IMMOBILIZATION IN CARBON NANOTUBES

SIMPLE ADSORPTION

Enzymes added to CNT functionalized with polymers combined with ionic liquids

Feng, W., & Ji, P. (2011). Enzymes immobilized on carbon nanotubes. Biotechnology advances, 29(6), 889-895.

Enzyme Lysozyme

Uses Recommended in treatment of certain ulcers, measles, multiple sclerosis, some skin diseases, and postoperative infections (antibacterial agent) Biosensor and articial kidneys Treatment of acatalasemia and removal of hydrogen peroxide

Enzyme Hyaluronidase

Uses Hydrolyses polyhyaluronic acid, a relatively impermeable polymer found between human cells; administered to increase diffusion of coinjected compounds. Removal of heparin after surgery Prevention and removal of blood clots Anti-inammatory agent

Urease Catalase

Heparinase Urokinase Streptodornase

G-6-P dehydrogenase Treatment of jaundice

Collagenase
Glucose Oxidase Asparaginase Trypsin

Skin ulcers
Glucose test in blood and urine Anticancer agent (leukemia) remove dead tissue from wounds, burns, and ulcers to speed the growth of new tissue and skin grafts, Anti inflammatory agent, dissolution of blood clots in

TPA
Tyrosinase Bilirubin oxidase Alkaline phosphatase

Dissolution of blood clots

Enzyme essential for the production of cellular pigments Treatment of neonatal jaundice Treatment of hypophosphatasia

Streptokinase

Ref: S. A. Costa, Helena S. Azevedo, and Rui L. Rei. CRC Press LLC .2005.Enzyme Immobilization in Biodegradable

Enzyme delivery
Nanospheres, nanocapsules, liposomes, micelles, and other nanoparticulates are frequently referred to as carriers for delivery of therapeutic and diagnostics agents. Asparagine is an essential amino acid for certain types of leukemias that lack asparaginase synthetase activity. Some success has been achieved in administering asparaginase in capsules made of nylon and polyurea to mice and rats. Many materials, such as nylon, polysulfone, glycidyl methacrylate, chitosan, cellulose, and cellulose derivatives, have been used for trypsin immobilization.

Problems and Solutions

The enzyme be recognized as foreign by the body and potentially severe immunogenic responses willbe stimulated, resulting in hypersensitivity reactions. To overcome these problems, immobilized enzyme derivatives have been prepared on various supports for extracorporeal treatment.

Extracorporeal treatment
Extracorporeal shunts have been proposed for the treatment of several clinical conditions. The most likely applications for enzymatic treatment are the removal of urea during kidney failure, removal of toxins (e.g., paracetamol) during liver failure, or the reduction of key metabolites from the circulation to treat cancer.

Urease Artificial Kidney

Urease is an enzyme that catalyzes the hydrolysis of urea to form ammonia and carbon dioxide. Hence, its immobilization by entrapment has been investigated by many workers for applications in biosensors and as articial kidneys. The most attention has been given to the development of enzyme reactors, where the urea would be removed and the dialysis uid prepared or further use. The use of this enzyme is often limited due to its high cost, availability in small amounts, instability, and the limited possibility of feasible recovery of these biocatalysts from a reaction mixture.

Inventors

Hugh D. Conlon, Robert S. Langer, Jr., Robert S. Lees, Claudy J. P. Mullon

Original Assignee

Massachusetts Institute Of Technology

A method for lowering the level of low density lipoprotein (LDL) in a subject's blood, which method comprises the steps of: a) immobilizing on a surface an enzyme selected from the group consisting of phospholipases A.sub.s ; b) contacting the LDL-containing whole blood or plasma with the immobilized enzyme to hydrolyze an ester bond at position 2 of glycerophospholipids present in LDL, thus modifying the LFL in a manner which allows the modified LDL to be more rapidly metabolized; and, c) circulating the modified LDL-containing whole blood or plasma in the subject's blood stream for ultimate metabolism of the modified LDL.

Red Blood Cell and Liposome as a Carrier for Enzyme

As a new therapeutic approach, human and animal red blood cells (RBCs) can been used as the carrier vehicle for a number of exogenous enzyme intended to be disseminated in an organism. Like RBCs, liposomes can also fulfill the promise of a drug-carrier vehicle because of their ability reduce toxicity and enhance efficacy of the agents that are encapsulated. However, therapeutic application of liposomes administered by the intravenous route has been limited, primarily because of a rapid clearance of liposomes from the bloodstream and their uptake by the macrophage cellsin the liver and spleen.

Thank You

Discussion
How can be RBCs used to immobilize enzymes? Is there difference while treating patients with immobilized enzymes and with treating with drugs in terms of time taken for clinical trials? Why was extracorporeal system no so successful? Are there systems which can be implanted inside the body? Can carbon nanotube immobilized enzymes be directly applied to the body? Are carbon nanotubes toxic to our body?

References
Feng, W., & Ji, P. (2011). Enzymes immobilized on carbon nanotubes. Biotechnology advances, 29(6), 889-895. S. A. Costa, Helena S. Azevedo, and Rui L. Rei. CRC Press LLC 2005.Enzyme Immobilization in Biodegradable Polymers for Biomedical Applications. 301-32 Hugh, D.C., Langer, R.S., Lees, R.S., Mullon, J.P.C., (1989) Patent Identifier No. US 5232696 A http://jackandraka.net/about.html