TY  - JOUR
AB  - Technical crystallization is an attractive method to purify recombinant proteins. However, it is rarely applied due to the limited crystallizability of many proteins. To overcome this limitation, single amino acid exchanges are rationally introduced to enhance intermolecular interactions at the crystal contacts of the industrially relevant biocatalyst Lactobacillus brevis alcohol dehydrogenase (LbADH). The wildtype (WT) and the best crystallizing and enzymatically active LbADH mutants K32A, D54F, Q126H, and T102E are produced with Escherichia coli and subsequently crystallized from cell lysate in stirred mL-crystallizers. Notwithstanding the high host cell protein (HCP) concentrations in the lysate, all mutants crystallize significantly faster than the WT. Combinations of mutations result in double mutants with faster crystallization kinetics than the respective single mutants, demonstrating a synergetic effect. The almost entire depletion of the soluble LbADH fraction at crystallization equilibrium is observed, proving high yields. The HCP concentration is reduced to below 0.5% after crystal dissolution and recrystallization, and thus a 100-fold HCP reduction is achieved after two successive crystallization steps. The combination of fast kinetics, high yields, and high target protein purity highlights the potential of crystal contact engineering to transform technical crystallization into an efficient protein capture and purification step in biotechnological downstream processes.
AU  - Grob, P.*
AU  - Huber, M.*
AU  - Walla, B.*
AU  - Hermann, J.*
AU  - Janowski, R.
AU  - Niessing, D.
AU  - Hekmat, D.*
AU  - Weuster-Botz, D.*
C1  - 58870
C2  - 48409
CY  - Postfach 101161, 69451 Weinheim, Germany
TI  - Crystal contact engineering enables efficient capture and purification of an oxidoreductase by technical crystallization.
JO  - Biotechnol. J.
VL  - 15
IS  - 11
PB  - Wiley-v C H Verlag Gmbh
PY  - 2020
SN  - 1860-6768
ER  - 

TY  - JOUR
AB  - Lung diseases belong to the major causes of death worldwide. Recent innovative methodological developments now allow more and more for the use of primary human tissue and cells to model such diseases. In this regard, the review covers bronchial air-liquid interface cultures, precision cut lung slices as well as ex vivo cultures of explanted peripheral lung tissue and de-/re-cellularization models. Diseases such as asthma or infections are discussed and an outlook on further areas for development is given. Overall, the progress in ex vivo modeling by using primary human material could make translational research activities more efficient by simultaneously fostering the mechanistic understanding of human lung diseases while reducing animal usage in biomedical research.
AU  - Zscheppang, K.*
AU  - Berg, J.V.*
AU  - Hedtrich, S.*
AU  - Verheyen, L.*
AU  - Wagner, D.E.
AU  - Suttorp, N.*
AU  - Hippenstiel, S.*
AU  - Hocke, A.C.*
C1  - 51789
C2  - 43494
CY  - Weinheim
TI  - Human pulmonary 3D models for translational research.
JO  - Biotechnol. J.
VL  - 13
IS  - 1
PB  - Wiley-v C H Verlag Gmbh
PY  - 2018
SN  - 1860-6768
ER  -