Molecular lyophilic

Gel Filtration. The lyophilized protein was redissolved in 50 mM phosphate buffer, pH 7.4 0.15 m NaCl 0.013 % sodium azide and loaded on a Superdex 75HR1030 column equilibrated with the same buffer. Elution was downward flow (0.15 ml/min) and 0.25 ml fi actions were collected. Fractions with pectin lyase activity were combined, dialyzed against distilled water and used in the next step. To estimate the molecular mass of PNL, the column was calibrated with standard proteins (Sigma MW-GF-70 Albumin, 66,000 Da Carbonic Anhidrase, 29,00 Cytochrome, 12,400 and Aprotinin, 6,500). The proteins were eluted in the conditions described above and their volumes (F ) were calculated fi om the peak maximum of the absorbance at 280 nm. The partition coefficient was obtained fi om the relationship where F, represents the bed volmne of column and F the void volume (which was calculated using blue dextran. Sigma). The molecular mass was determined using a standard curve of vs the logarithm of the molecular masses of the standards [28, 29]... 

Exudate collection in trap solutions usually requires subsequent concentration steps (vacuum evaporation, lyophilization) due to the low concentration of exudate compounds. Depending on the composition of the trap solution, the reduction of sample volume can lead to high salt concentrations, which may interfere with subsequent analysis or may even cause irreversible precipitation of certain exudate compounds (e.g., Ca-citrate, Ca-oxalate, proteins). Therefore, if possible, removal of interfering salts by use of ion exchange resins prior to sample concentration is recommended. Alternatively, solid-phase extraction techniques may be employed for enrichment of exudate compounds from the diluted trap solution (11,22). High-molecular-weight compounds may be concentrated by precipitation with organic solvents [methanol, ethanol, acetone 80% (v/v) for polysaccharides and proteins] or acidification [trichloroacetic acid 10% (w/v), per-... 

Fractionations. Following lyophilization of the polar leachate fractions, separations by exclusion chromatography suggest three major molecular weight regions, labelled peak 1A, peak 1B and peak 2 (Figure 3). Retention times on a G—15 Sephadex gel indicate molecular weights of peaks 1A and 1B compounds to be between 600 and 1000. 

The reaction solution was diluted with 36 mL of water. The unreacted free drug and other low molecular weight materials were removed by a Centriprep-10 concentrator. Purification was repeated until HPLC analysis (Bio-Sil TSK-250) of the product indicated the absence of free drug. The final purified product was lyophilized to afford 483 mg of a yellow solid. This solid product was readily soluble in water or aqueous buffer. The amount of drug covalently bound to MVE was estimated by the absorbance at 303 nm using a molar extinction coefficient of 17.03 x 103. The MVE-y-hy-MTX contains 26% methotrexate-y-hydrazide by weight. 

After cooling to room temperature, the sugar-dendrimer derivative may be precipitated with a large volume of methanol. The precipitate may be purified from reaction products by dialysis against water or buffer using a membrane with a molecular weight cutoff of 3500 Daltons. The final product may be stored frozen or lyophilized to a white powder. 

Yoshika, S., Asu, Y., Kojima, Sh. Determination of molecular mobility of lyophilized bovine serum albumin and gamma-globulin by solid-state H1 NMR and relation to aggregation-susceptibility. Pharm. Res. 13 (6), p. 926-930, 1996... 

Molecular imprinting is not limited to organic polymer matrices, but can also be applied to silica-based materials and even proteins. Proteins freeze-dried in the presence of a transition state analogue as template have been used successfully as catalysts, e.g., for the dehydrofluorination of a fluorobutanone. For instance, lyophilized 3-lactoglobulin imprinted in this manner with N-isopropyl-N-ni-trobenzyl-amine could accelerate the dehydrofluorination by a factor of 3.27 compared to the non-imprinted protein see Table 5 [62]. In a similar procedure, BSA was imprinted with N-methyl-N-(4-nitrobenzyl)-S-aminovaleric acid and showed an enhancement of the catalytic effect by a factor of 3.3 compared to the control protein for the same reaction see Table 5 [113]. 

Micelle solutions of PlPAAm-Ci8H35 was prepared by direct dissolution of the polymer in cold water (4°C) due to its good water solubility [23]. Each solution of PIPAAm-PSt, PlPAAm-PBMA, and PIPAAm-PLA was prepared by dissolving each copolymer in DMF, A-ethylacetamide, and DMAc, respectively. The solutions were put into a dialysis bag (MWCO = 13,000) and dialyzed against distilled water at 10°C, 20°C, and 4°C, respectively, for 24 hours. The micelles were purified with ultrafiltration membrane of 200,000 molecular weight cut off at 4°C. The aqueous solution was lyophilized to leave a white powder of micelles. 

Dialysis and ultraflltration have been largely applied to isolate and fractionate food proteins and peptides. To isolate the protein fraction from wine and must samples, different authors used dialysis followed by lyophilization to concentrate the dialyzed samples [106,108,109], Depending on the application, membrane of different material, filtration surface and cut-off, able to fractionate the molecules in function of their molecular size, can be used to remove either proteins and other macromolecules or amino acids and small peptides. 

The powdered roots of C. tinctorium were extracted with ethanol (% 96, v/v) using a soxhiet apparatus to remove low molecular weight compounds. Extraction procedures continue until no color could be observed in the ethanol. The residue was extracted with water at 50 °C, 2 hour for two times. Obtained extract was filtered through gauze and Whatman GF/A glass fiber filter and then concentrated at 40 °C in vacuum and dialysed at cut-off 3500 Da to give a 50 °C crude extract. The extracts was kept at-18 °C or lyophilized. 

The newer supports for gel chromatography, such as lyophilized polystyrene14 and porous silica,21-23 have been successfully applied to the fractionation of dextrans over a wide range of molecular weight. 

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