Purification streamlining

STREAMLINE expanded bed adsorption is particularly suited for large scale recombinant protein and monoclonal antibody purification. STREAMLINE adsorbents are specially designed for use in STREAMLINE columns. The technique requires no sample clean up and so combines sample preparation and capture in a single step. As shown in Figure 10, crude sample is applied to an expanded bed of STREAMLINE media, target proteins are captured whilst cell debris, particulate matter, whole cells, and contaminants pass through. Flow is reversed and the target protein is desorbed in the elution buffer. 

Artolozaga, M. J., Jonas, R., Schneider, A. L., Furlan, S. A., and Carvalho-Jones, M. F., One step partial purification of fi-D-ga lactosidase from Kluyveromyces marxianus CDB 002 using STREAMLINE-DEAE, Bioseparation, 7, 137, 1998. 

The assembly of tetrapeptide 19 that contains all possible 0-dipeptide bonds, (03-03)-, (03-02)-, and (02-03), and also a turn inducing 03-(R)-Ala-02-(R)-Val element was achieved employing a Boc-strategy (Scheme 5). A fluorous benzyl group was incorporated in the first amino acid to streamline the purification procedure by fluorous solid phase extraction (LSPE) (Lilippov et al. 2002 de Visser et al. 2003). Thus, the assembly of the fully protected tetrapeptide commenced with the construction of the first 03-03-peptide bond by applying the previously established conditions. A residence time of 3 min at 90°C provided the Boc-protected dipeptide 15 in 91% isolated yield after LSPE. Notably, the product precipitated in the collection flask, which was kept at ambient temperature, indicating the poor solubility of this class of compounds (Hessel et al. 2005). 

An attractive feature of this dehydrative coupling approach is that it avoids the need for isolation of intermediate glycosyl donors. This can be desirable if a glycosyl donor is not stable to isolation or purification. Moreover, the use of a hemiacetal donor reduces the number of synthetic manipulations of the carbohydrate donor by avoiding hemiacetal derivatization to alternative donor types. In this way, the approach has the potential to streamline time and labor-intensive multiglycosylation sequences. Although there increasingly have been reports of these direct dehydrative... 

His tag. The ability of the imidazole moiety of the histidine residue to bind divalent metal ions such as nickel, iron, and cobalt can be used to purify histidine-containing proteins on a column which has such divalent metals bound to it. To streamline the purification procedure of any desired protein, such histidines are deliberately added to a protein of choice by cloning a four- to sixfold CAG repeat sequence into the expression construct upstream of the gene of interest so as to express an N-terminal or C-terminal tetra- or hexa-His tag. From experience, especially in E. coli, C-terminal tagging often yields superior results to N-termi-... 

Antibody capture of viruses can be used as a preparatory step in nucleic acid amplification techniques. Immunocapture of virus particles can be used to streamline and/or optimize the concentration, purification and specificity requirements of polymerase chain reaction assays. 

Chono, H. (1998). Purification of recombinant proteins from E. coli using Streamline, lnt. Conf. Expanded Bed Adsorption, Napa Valley, CA, 1998, Abstr., p. 9.3. 

Noronha, S., Kaufman, J., and Shiloach, J. (1999). Use of STREAMLINE Chelating for capture and purification of poly His Tagged recombinant proteins. Bioseparation 8, 145-151. 

Thus, the selective enrichment of the target analyte was successfully demonstrated using the imprinted polymer. Conventional SPE sometimes needs to be combined with a different type of SPE or other separation steps to complete pre-purification, because compounds with similar chemical properties may accompany the analyte as impurities. On the other hand, imprinted polymer is an affinity-type SPE sorbent that exhibits specificity for an analyte therefore, the imprinted polymer-based SPE is able to streamline the whole procedure of analysis. Although aqueous conditions were employed here, it is also notable that the utility in organic solvents is one of the useful characteristics of imprinted polymers as SPE sorbents [18,19]. 

O. Bertrand, S. Cochet, and J. P. Cartron, Expanded bed chromatography for one-step purification of mannose binding lectin from tuhp bulbs using mannose im-mobihzed on DEAE Streamline, J. Chromatogr. A 822 19 (1998). 

EBA is a single pass operation in which target proteins are purified from crude sample, without the need for separate clarification, concentration and initial purification to remove particulate matter. Crude sample is applied to an expanded bed of STREAMLINE adsorbent particles within a specifically designed STREAMLINE column. Target proteins are captured on the adsorbent. 

Purification protamine sulfate and DEAE-Streamline, HIC (Phenylsepharose), protamine sulfate and IEX (DEAE-Toyopearl)... 

There is always a need for streamlining the initial purification steps of products that are derived from fermentation. Recent advances in a technique called expanded-bed adsoiption have shown the ability to elimmate the solid-4iquid separation step and thereby increase product yield and reduce processing time. Expanded-bed adsorption is not a new concept, since there are reported applications for streptomycin (23) in the 1950s and novobiocin (24) in the 1970s. However, these applications used adsorbents that were not specifically designed for expanded-bed-adsorption procedures and often required compli-... 

Most reported applications have been for protein purification (28-30). One example (31) was for direct recovery of Annexin V, an intracellular recombinant protein, from E. coli homogenate, using the Streamline DEAE adsorbent (Pharmacia). The procedure was developed in a laboratory-scale column (50 mm diameter by 1000 mm length) and then successfully scaled to a pilot-plant column (200 mm diameter by 950 mm length). The yield was 95% for both laboratory and pilot-plant runs and achieved a threefold reduction in volume compared to the homogenate. 

The quinoline scaffold and derivatives occur in a large number of natural products and drug-like compounds. A method for microwave-assisted synthesis of 2-aminoquinolines has been described by Wilson et al. [62]. The process involves rapid microwave irradiation of secondary amines and aldehydes to form enamines, then addition of 2-azidobenzophenones with subsequent irradiation to produce the 2-aminoquinoline derivatives (Scheme 10.27). Purification of the products was accomplished in a streamlined manner by using solid-phase extraction techniques to produce the desired compounds in high yields and purity. Direct comparison of the reaction under thermal and microwave conditions, using identical stoichiometry and sealed reaction vessels, showed the latter resulted in improved yield. 

Noronha S., Kaufman J. and Shiloach J. 1999. Use of streamline chelating for capture and purification of poly-His-tagged recombinant proteins. Bioseparation, 8, 145. 

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