Deep-well plates

A combinatorial approach for biocatalytic production of polyesters was demonstrated. A library of polyesters were synthesized in 96 deep-well plates from a combination of divinyl esters and glycols with lipases of different origin. In this screening, lipase CA was confirmed to be the most active biocatalyst for the polyester production. As acyl acceptor, 2,2,2-trifluoroethyl esters and vinyl esters were examined and the former produced the polymer of higher molecular weight. Various monomers such as carbohydrates, nucleic acids, and a natural steroid diol were used as acyl acceptor. 

Fig. 4.6 Ethos combCHEM system (Milestone, Inc.) left 96 deep-well plates right overhead rotor with two plates. Operating limits 200 °C, 5 bar.

Hydrogen evolution assay in deep well plates... 

Figure 4. Photograph of a two ml deep well plate with MicroMap used for the mutant screening.

The photograph shows the two ml standard deep well plate with 1.6 ml TAP-agar. Transformant colonies where picked from plate, spread inside the wells with plating-beads and incubated for 12 days at 20°C under illumination. The plate was afterwards sealed with a MicroMat ... 

There are two yeast expression hosts that have an established track record for high-level production of heterologous proteins, namely Saccharomyces cerevisiae and Pichia pastoris. HTP expression screening using microplate formats has been reported for both these yeasts by Lang and coworkers (Holz et ah, 2002, 2003 Boettner et ah, 2002). In both cases standard protocols have been miniaturized with cells cultured in either 1.5 ml cultures in 96-deep-well plates for S. cerevisiae or 2 ml cultures in 24-deep-well plates for P. pastoris. Soluble... 

MASTERBLOCK, 96-well, 2 ml, V-bottom (Greiner Bio-One, Cat. No. 780270). This product is a deep well plate in a 96-well format to culture E. coli. Other equivalent 96-well plates can be used. One should select a product that can be repeatedly used by autoclaving for saving costs. 

Pick up bacterial cells from the LB agar plate by wide side of teeth pick and rinse them into 1 ml of LBG medium in 96-deep-well plate covered with an AirPore Tape Sheet. 

All bacteria were cultivated in a 96-deep-well plate, and their ATP synthetic activities were assayed using a 96-well format. Cellular ATP synthetic activity of C. lutamicum was highest among the bacteria tested. Activities were the mean ( = 3)... 

Using a liquid handler, transfer the contents of each test tube to its corresponding well in a 2-mL 96-well polypropylene deep-well plate for purification by HPLC. 

Using a liquid handler, remove 5 iL of the solution from each well, dilute the aliquot to 1.0 mL with MeOH/water 95 5 in a new deep-well plate for LC/MS analysis. 

The reverse phase HTP separations of aqueous extracts are highly reproducible. The aqueous extracts from whole plant of Ainsliaea henryi were separated twelve times on 4 parallel C-18 columns and a total of twelve 96-deep well plates generated. The HTP/UV chromatograms from 12 column separations were identical and the samples were combined based on the same well position from the twelve plates (data not shown). 

In order to identify the active compounds from the plant extracts, a high throughput fractionation process was executed. The active organic and aqueous extracts were fractionated using two different HTP methodologies as illustrated in previous section. The fractions were collected as described into a 96 deep-well plate. Each of the fractions was tested for its ability to inhibit... 

Comer et al. developed a high-throughput pKa assay (SGA profiler, Sirius Analytical) which is able to measure the pKas of up to 200 compounds between pH2 and pH 12. Compounds are dissolved in DMSO to give 10 mM stock solutions. 5-20 xl of the stock solution is transferred into a deep well plate and is... 

An example for an automated stability test in plasma is described by Linget and du Vignaud (1999). Incubations are performed on a 215 Gilson liquid handler. Incubation was done at substrate concentrations of 50 pM on 96 deep well plates. Each incubation tube contained 375 pL of a 200 pM test compound solution (in 0.1 M Tris buffer with 3% BSA, added to assist dissolution of compounds with poor solubility) and 1125 pL of plasma. Samples are taken after incubation times of 0, 1, 2, 3, 4 and 5 min. At each of these time points an aliquot of the incubation mixture was transferred from the incubation tube into a well in a 96 deep well plate containing an equal volume of acetonitrile for quenching by protein precipitation followed by centrifugation of the plates. Supernatants were analyzed by HPLC for metabolic screening. 

The Affymax Research Institute has developed an original approach for the synthesis of organic compounds in 96 deep well plates. The high-throughput organic parallel synthesis (HiTOPS) systemP l (Figure 4) utilizes a variety of 96 deep well filtration microtiter plates available from Whatman Polyfiltronics. 

Another separation method involves cooling of the organic/aqueous phase to -20 °C in deep-well plates in the presence of pins. After the freezing process, the aqueous phase can be removed as ice attached to the array of pins while the organic phase remains in the deep-well plate. By this so-called lollipop method, 96 aqueous/organic mixtures can be easily separated [12]. 

To each row in a deep-well microtiter plate were added a 1.5 m solution (0.4 mL for each well) of each nitroalkane, a 1.5 m solution (0.4 mL for each well) of phenyl isocyanate, and a 0.0225 m solution (0.4 mL for each well) of EtsN in dioxane. Alkene-functionalized pins (213) secured in a pin holder block were put into each deep-well microtiter plate containing the above solution, and the assembled set (pin holder plus deep-well plate) was placed in a stainless steel container with the lid tightly sealed, where the reaction mixture was incubated overnight at 60 °C. The pins, secured in a pin holder block, were washed with dioxane and DMF in a chemically resistant polypropylene bath and dried. The [2 -i- 3]-cycloaddition reaction was repeated twice more (as above but for 6 h). Finally, pins in a pin holder block were washed with dioxane, DMSO (60 °C), DMF, THF, and DCM (250 mL for each solvent) in a chemically resistant polypropylene bath and dried to give isoxazoline-functionalized pins (214). 

---