Developing effective therapies against chronic wound healing deficiencies is a global priority. Thus we evaluated the safety of two different doses of topically administered autologous APOSEC, the secretome of apoptotic peripheral blood mononuclear cells (PBMCs), in healthy male volunteers with artificial dermal wounds.
Ten healthy men were enrolled in a single-center, randomized, double-blinded, placebo-controlled phase 1 trial. Two artificial wounds at the upper arm were generated using a 4-mm punch biopsy.
Each participant was treated with both topically applied APOSEC and placebo in NuGel for 7 consecutive days. The volunteers were randomized into two groups: a low-dose group
(A) receiving the supernatant of 12.5 × 106 PBMCs and a high-dose group
(B) receiving an equivalent of 25 × 106 PBMCs resuspended in NuGel Hydrogel.
Irradiated medium served as placebo.
The primary outcome was the tolerability of the topical application of APOSEC.
All adverse events were recorded until 17 days after the biopsy. Local tolerability assessment was measured on a 4-point scale. Secondary outcomes were wound closure and epithelization at day 7.
No therapy-related serious adverse events occurred in any of the participants, and both low- and high-dose treatments were well tolerated. Wound closure was not affected by APOSEC therapy.
Contact sensitization to modern wound dressings in 70 patients with chronic leg ulcers.
Patients with chronic leg ulcers typically experience contact allergy to topical treatments. Although declared as hypoallergenic, modern wound dressings have caused several reported cases of contact allergy.
The aim of the study was to evaluate any allergenic potential of modern wound dressings in patients with leg ulcers.
Seventy-one patients were included in our prospective observation. Patch tests were performed with a selection of 10 modern wound dressings and with selected allergens according to series of the German Contact Dermatitis Research Group (DKG).
Of 70 patients eligible for evaluation, 12 (17%) were positive for the hydrogel NuGel, followed by the hydrocolloid NuDerm (n = 7/70, 10%) and the ionic silver-containing wound dressing Aquacel Ag and the gauze Adaptic (both n = 4/70, 5%).
Patients with recalcitrant ulcers of prolonged duration showed a significant higher number of epicutaneous sensitizations to wound dressings than patients with shorter ulcer duration.
The allergenic potential of modern wound dressings should not be underestimated. There is need for precise declaration of all ingredients.
Affinity purification and characterization of an anti-PEG IgM.
Anti-PEG IgM was purified by affinity chromatography using variable length PEG chains (5, 10, 20 and 30 kDa) as affinity ligands. Maximal binding of anti-PEG IgM was observed using the 30 kDa PEG-derivatized NuGel (single passage).
Purified anti-PEG IgM was characterized for binding to PEG functionalized proteins/peptides by surface plasmon resonance, western blotting and ELISA.
Anti-PEG IgM, in solution and adsorbed on 20 kDa PEG-derivatized NuGel, was subjected to pepsin digestion followed by affinity chromatography. SDS-PAGE analysis of eluates in both preparations yielded one fragment that was similar in size.
However, an additional lower molecular weight band was observed in solution-digested affinity purified material that was not present in the eluate from the material subjected to pepsin digestion on the affinity matrix.
The lower MW fragment could be eluted under milder conditions, suggesting loss of binding multiplicity. Analysis by mass spectrometry yielded molecular weights of 132 kDa (both) and 82 kDa (solution) for the respective fragments.
N-terminal sequencing of both fragments resulted in primary sequences (heavy and light chains) that were not only identical to each other but also to those of native IgM.
The anti-PEG IgM fragments were characterized for binding to pegylated interferon alfa-2a by ELISA. The results from these studies suggest that affinity purified anti-PEG IgM and fragments can be used as probes in detection assays for PEG functionalized biotherapeutics in pre-clinical and clinical studies.
Automated high-performance immunosorbent assay for recombinant leukocyte A interferon.
- A rapid assay for recombinant leukocyte A interferon has been developed that includes a small immunosorbent column (Amicon glass column ca. 10 X 6 mm; i.e., ca. 0.5 ml), containing monoclonal antibody, immobilized on Nugel-polyhydroxy phase silica, 500 A, 200-400 mesh (Diagnostic Specialties, Metuchen, NJ, U.S.A.).
- The column has been automated so that the operator need only inject sample (0.25 ml) every 18 min (or one can use an automatic sample injector) and initiate the program cycle of the microprocessor.
- A hard-copy result from an integrator is available in less than 20 min. Routine analyses were performed at a flow-rate of 4 ml/min in the concentration range 0.02-0.3 mg/ml. Reproducibility of the assay was checked by assaying the same crude extract seven times in succession. Standard deviation was 3.96% and correlation coefficient was 0.9996.
- The advantages of this technique include rapid analysis time and relative simplicity, compared to the enzyme immunoassay.
Isolation of proteins from crude mixtures with silica and silica-based adsorbents.
Silica and silica-based adsorbents have been used to isolate specific proteins from crude mixtures. Acid-activated Nugel silica and Nugel SP-Silica were used to extract human immune interferon from the conditioned medium of mixed lymphocyte cultures.
NuGel-BindPro™ |
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| BPM55-15 | Biotech Support Group | 15 preps | 540 EUR |
NuGel-BindPro™ |
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| BPM55-50 | Biotech Support Group | 50 preps | 1076.4 EUR |
NuGel™ Poly-Amine |
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| NPAM-25 | Biotech Support Group | 25 Grams | 559.2 EUR |
NuGel™ Poly-Aldehyde |
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| NPAY-25 | Biotech Support Group | 25 Grams | 559.2 EUR |
NuGel™ Poly-Carboxy |
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| NPCY-25 | Biotech Support Group | 25 Grams | 559.2 EUR |
NuGel™ Poly-Epoxy |
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| NPEY-25 | Biotech Support Group | 25 Grams | 559.2 EUR |
NuGel™ Poly-Hydroxy |
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| NPHX-25 | Biotech Support Group | 25 Grams | 559.2 EUR |
NuGel™ Phenyl Boronic Acid |
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| NPBA-05 | Biotech Support Group | 5g | 814.8 EUR |
NuGel™ Phenyl Boronic Acid |
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| NPBA-10 | Biotech Support Group | 10g | 1143.6 EUR |
NuGel™ Glycoprotein Enrichment PBA Kit |
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| NGPBA-10 | Biotech Support Group | 10 preps | 571.2 EUR |
NuGel™ Glycoprotein Enrichment PBA Kit |
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| NGPBA-50 | Biotech Support Group | 50 preps | 2032.8 EUR |
NuGel-HemogloBind™ (Hemoglobin Removal) Kit |
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| NP-HO-T25 | Biotech Support Group | 25 preps | 771.6 EUR |
NuGel-HemogloBind™ (Hemoglobin Removal) Kit |
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| NP-HO-T50 | Biotech Support Group | 50 preps | 1411.2 EUR |
Human interleukin-2 was extracted from the conditioned medium of Jurkat cells with LiChroprep RP-8. In each case, batch absorption was accomplished by gentle stirring in a microcarrier vessel.
The adsorbate was collected by sedimentation, and either batch-eluted or packed into a column for gradient elution. A high degree of concentration and purification was achieved with a high recovery of biological activity.