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Xu H.,New York University | Scott G.M.,New York University | Jiang F.,Cell Genesys Inc. | Kelly C.,Oregon State University

Manganese peroxidase (MnP) is the main enzyme implicated in the biobleaching of kraft pulps by white-rot fungi. However, potential commercial applications of this enzyme have been limited by its availability in large quantities. Advances have been made to produce high-yield concentrated recombinant MnP (rMnP). The objective of this study was to evaluate the ability of rMnP to delignify and brighten kraft pulps. The rMnP, produced from the yeast Pichia pastoris - in high-cell density and in fed-batch fermentations - was found to be effective in lignin removal in both hardwood and softwood unbleached kraft pulps. The rMnP applied at 30?U g -1 pulp for 24?h followed by alkali extraction caused significant kappa number reductions for all the pulps tested with different initial lignin contents and structures. Softwood and hardwood pulps showed similar delignification rates during rMnP treatments. Highly delignified pulps with kappa number less than 10 are less susceptible to delignification by rMnP compared with the pulps with higher lignin content. The rMnP-treated pulp was also shown to be more susceptible to subsequent peroxide bleaching compared with the control pulp. More than 60% of the kappa number reduction was achieved by sequential rMnP treatments combined with alkaline extraction. Sequential treatment with xylanase and rMnP also resulted in more extensive delignification than in each enzyme treatment alone or in the case of simultaneous application of the enzymes. © 2010 by Walter de Gruyter Berlin New York. Source

Cell Genesys Inc. | Date: 2006-11-07

Pharmaceuticals for immunotherapy and for the treatment of cancer, prostate diseases, pancreatic diseases, and leukemia.

Huang L.-C.,Cell Genesys Inc. | Huang L.-C.,Genentech | Lin W.,Cell Genesys Inc. | Yagami M.,Cell Genesys Inc. | And 5 more authors.

A method using Cedex automatic cell counter (Innovatis) to determine the cell density and viability of a whole cell-based immunotherapy product has been developed and validated for the assay performance characteristics including specificity, accuracy, precision, linearity, range, and robustness. Instrument-to-instrument variation due to intrinsic differences in handmade flow cells was also evaluated. For cell density, Cedex demonstrated acceptable specificity, accuracy and precision for cell densities ranging from 3.13×105 to approximately 1.0×107cells/mL, with intermediate precision of about 5% relative standard deviation (RSD). However, a marked difference was observed between the two instruments studied and they therefore could not be used interchangeably without additional calibration procedures that went beyond the manufacturer's recommendation. For viability, mixing known numbers of non-viable cells with highly viable cells allowed evaluation of the specificity, accuracy and linearity of the viability determination. Acceptable levels of accuracy (95.3-106.4% recovery) and precision (RSD<5%) were demonstrated for the viability range from 50 to 100%. The instrument-to-instrument difference was less than 4.6%. The assays for both cell density and viability were sufficiently robust for assay parameters. However, the effect of certain parameters was cell line-dependent, suggesting that Cedex performance should be verified for each cell line of interest. © 2010 The International Association for Biologicals. Source

Liang S.C.,Cell Genesys Inc. | Liang S.C.,Nodality | Moskalenko M.,Cell Genesys Inc. | Moskalenko M.,Genentech | And 3 more authors.
Clinical Immunology

In this report, a Treg-depleting anti-FR4 antibody is combined with a GM-CSF-secreting tumor cell immunotherapy (GVAX) for treatment of melanoma-bearing animals. Median survival time (MST) of animals treated with GVAX was 41. days, compared to a MST of 32. days in untreated animals. Anti-FR4 monotherapy had no effect on MST. Combination of anti-FR4 and GVAX significantly prolonged MST to 55. days, suggesting that these two agents can function cooperatively. Combination therapy increased expression of IFN-γ and granzyme B by CD8 T cells. In contrast to anti-CD25-mediated Treg depletion, administration of anti-FR4 after GVAX did not reduce efficacy, suggesting that anti-FR4 does not deplete effector cells induced by GVAX. Triple combination of a blocking CTLA4 antibody with GVAX and anti-FR4 further enhanced overall survival and reduced growth of well-established melanomas. Considered together, anti-FR4 antibody and GVAX may be a promising approach for the treatment of patients with cancer. © 2013 Elsevier Inc. Source

Burke J.M.,Billings Clinic | Lamm D.L.,University of Arizona | Meng M.V.,University of California at San Francisco | Nemunaitis J.J.,Mary Crowley Medical Research Center | And 8 more authors.
Journal of Urology

Purpose: We assessed the safety, pharmacokinetics and anticancer activity of intravesical CG0070, a cancer selective, replication competent adenovirus, for the treatment of nonmuscle invasive bladder cancer. Materials and Methods: A total of 35 patients received single or multiple (every 28 days × 3 or weekly × 6) intravesical infusions of CG0070 at 1 of 4 dose levels (1 × 1012, 3 × 1012, 1 × 1013 or 3 × 1013 viral particles). Response to treatment was based on cystoscopic assessment and biopsy or urine cytology. Urine and plasma CG0070, and granulocyte-monocyte colony-stimulating factor were measured in all patients. A subset of 18 patients was assessed for retinoblastoma phosphorylation status. Results: Grade 1-2 bladder toxicities were the most common adverse events observed. A maximum tolerated dose was not reached. High levels of granulocyte-monocyte colony-stimulating factor were detected in urine after administration in all patients. Virus replication was suggested based on an increase in urine CG0070 genomes between days 2 and 5 in 58.3% of tested patients (7 of 12). The complete response rate and median duration of the complete response across cohorts was 48.6% and 10.4 months, respectively. In the multidose cohorts the complete response rate for the combined groups (every 28 days and weekly × 6) was 63.6% (14 of 22 patients). In an exploratory, retrospective assessment patients with borderline or high retinoblastoma phosphorylation who received the multidose schedules had an 81.8% complete response rate (9 of 11). Conclusions: Intravesical CG0070 was associated with a tolerable safety profile and antibladder cancer activity. Granulocyte-monocyte colony-stimulating factor transgene expression and CG0070 replication were also suggested. © 2012 American Urological Association Education and Research, Inc. Source

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