Synthetic Methodology for Asymmetric Ferrocene Derived Bio-conjugate Systems via Solid Phase Resin-based Methodology

The overall goal of the following experiment is to produce asymmetric ferne, biotin, bio conjugates that can be immobilized on a gold surface. This is achieved by synthesis of the ferne biotin bio conjugate on a solid phase resin. As a second step, the bio conjugate is removed from the resin, which allows for isolation and characterization of the compound.

Next, the bio conjugate is incubated with a gold surface in order to allow adhesion of the thalates with the gold. The results show that a ferne bio conjugate can be produced in good yield and purity and then immobilized on a gold surface. The main advantage of using this technique over existing methods like solution-based methods, is that purification is simple and straightforward.

In addition to avoiding byproducts produced using solution based methods, So we first had the idea of this method when we thought about the efficiency and practicality of the glucose sensors that are used to monitor diabetic patients. Demonstrating this procedure will be Sean Rodich from my lab and Paulina Gonzalez undergraduate and graduate students respectively from my laboratory. First place, 250 milligrams of biotin loaded resin into a fritted syringe.

Swell the resin by drawing up five milliliters of dimethylformamide and shaking the syringe on a lab shaker for 20 minutes. When finished, expel the solution and repeat the dimethylformamide swelling. Following this, remove the FOC protecting group by adding four to six milliliters of 20%papine in dimethylformamide to the syringe, followed by 10 to 15 minutes of shaking After repeating the deep protection process, wash the resin three times with a sequence of dimethylformamide, one-to-one dimethylformamide and methanol, one-to-one methanol and di chloro methane and di chloro methane.

After expelling the solution, perform a hyn test on a small sampling of the beads to confirm successful deep protection by the presence of blue upon heating. Next, mix a solution containing one prime FM amino farin, one carboxylic acid, HOBT hydrate, DIC di, isopropyl ethyl amine, and a four to one mixture of di chloro methane and dimethylformamide. Draw this solution into the fritted syringe and gently shake on a lab shaker for six hours.

After washing the resin, perform the hyran test as previously described to confirm that coupling has occurred. Following this, remove the FM o group by the addition of 20%ine in dimethylformamide to the resin in the syringe after shaking wash in the same manner as before. Prepare a solution composed of fmm cyst trit till O-H-O-B-T hydrate, DIC di, isopropyl eth lamine, and a four to one mixture of di chloro methane and dimethylformamide.

Add this cystine coupling cocktail to the fritted syringe and gently shake for six hours. After confirming the coupling using the HYN test, remove the F MOC component with 20%ine. Once washing is complete, verify the free terminal amine using the hydrant test.

At this point, make a solution of trichloroacetic acid water one, two, et ethane ol, and try isopropyl cy. Then add the solution to the syringe. After shaking for four hours, collect the resulting red brown solution in an einor tube and evaporate the trichloroacetic acid slowly using a stream of air.

Following this at approximately 15 milliliters of cold dathyl ether to the einor tube to precipitate the product, which will form with gentle agitation. Isolate the product via centrifugation. Then repeat cycles of d ethyl, ether, washings, and centrifugation.

To obtain the ferne biotin bio conjugate as a red brown solid. Confirm the identity of the ferne biotin bio conjugate using proton and carbon NMR spectroscopy and E-S-I-M-S analysis. Perform HPLC and elemental analysis to confirm the composition of the isolated compound.

Once the product has been fully characterized, cut polymer backed gold slices into squares of approximately 0.25 square inches. Then fill a 50 milliliter beaker with a one millimolar deionized water solution of the ferne biotin bio conjugate. Add one of the gold slides to the beaker and cover with a watch glass.

Allow the slide to incubate overnight at room temperature with no agitation. After incubation, remove the gold slide from the solution and wash with deionized water. Once the slide is dry, obtain scanning electron microscopy images using a scanning electron microscope to observe the immobilized ferne biotin bio conjugate.

The resin bound form of ferne biotin bio conjugate one is shown here. The covalent attachment of the ferne component gives rise to an orange tint to the resin beads that is indicative of an immobilized iron containing complex as opposed to iron absorption by the PEG component of the resin bead. Following removal of the compound from the resin beads, the resulting purity and yield resulting is far superior to typical solution methodology.

Once mastered, this sign can be performed in three days if performed properly. Don't forget that working with chlor acidic acid and F and diet can be extremely hazardous precautions as using personal protective equipment and carrying out the procedure inside the fume Hood should always be taken when performing this procedure.