Application of platform process development approaches to the manufacturing of Mabcalin™ bispecifics.

Bispecific biotherapeutics offer potent and highly specific treatment options in oncology and immuno-oncology. However, many bispecific formats are prone to high levels of aggregation and instability, leading to prolonged development timelines, inefficient manufacturing, and high costs. The novel class of Mabcalin™ molecules consist of Anticalin® proteins fused to an IgG and are currently being evaluated in pre-clinical and clinical studies.

Metallothionein, essential elements and lipid peroxidation in mercury-exposed suckling rats pretreated with selenium.

Detoxification of mercury (Hg) with selenium (Se) in the early postnatal period with regard to the expression of metallothionein protein (MT), essential element status, and lipid peroxidation level in tissues has not been studied. Seven-day-old Wistar pups were orally pretreated with Se [6 μmol Na2SeO3/kg body weight (b.w.

Retrocyte Display® technology: generation and screening of a high diversity cellular antibody library.

Over the last nearly three decades in vitro display technologies have played an important role in the discovery and optimization of antibodies and other proteins for therapeutic applications. Here we describe the use of retroviral expression technology for the display of full-length IgG on B lineage cells in vitro with a hallmark of a tight and stable genotype to phenotype coupling. We describe the creation of a high-diversity (>1.

Redirecting catalysis from proteolysis to perhydrolysis in subtilisin Carlsberg.

Enzyme promiscuity describes the ability of biocatalysts to catalyze conversions beyond their natural reactions. Enzyme engineering to promote side reactions is attractive for synthetic and industrial applications. For instance, a subtilisin Carlsberg protease variant (T58A/L216W) catalyzes in addition to its proteolytic activity the generation of peroxycarboxylic acids from corresponding esters in the presence of hydrogen peroxide.

Mercury disposition in suckling rats: comparative assessment following parenteral exposure to thiomersal and mercuric chloride.

Due to the facts that thiomersal-containing vaccine is still in use in many developing countries, and all forms of mercury have recognised neurotoxic, nephrotoxic, and other toxic effects, studies on disposition of ethylmercury and other mercury forms are still justified, especially at young age. Our investigation aimed at comparing mercury distribution and rate of excretion in the early period of life following exposure to either thiomersal (TM) or mercuric chloride (HgCl₂) in suckling rats. Three experimental groups were studied: control, TM, and HgCl₂, with 12 to18 pups in each.

Vanadium bromoperoxidase-coupled fluorescent assay for flow cytometry sorting of glucose oxidase gene libraries in double emulsions.

A Vanadium bromoPeroxidase-coupled fluorescent assay (ViPer) for ultrahigh-throughput screening of glucose oxidase (GOx) gene libraries employing double emulsions and flow cytometry was developed. The assay is based on detection of the product of a GOx reaction, hydrogen peroxide, that is first converted to a hypobromide by vanadium bromoperoxidase in the presence of sodium bromide. The hypobromide is afterwards detected in a reaction with a fluorogenic probe, 3-carboxy-7-(4'-aminophenoxy)-coumarine, where fluorescent 3-carboxy-coumarine is released.

Salivary concentrations of nickel and chromium in patients with burning mouth syndrome.

It has been documented in vitro and in vivo that metal dental appliances release metal ions due to corrosion. Dentists must choose among many dental casting alloys available, often without knowledge of their biological properties and effect on oral mucosa. The aim of this study was to measure metal content of nickel (Ni) and chromium (Cr) in whole saliva of 85 patients with and without metal dental appliances.

Effect of selenium pre-treatment on antioxidative enzymes and lipid peroxidation in Cd-exposed suckling rats.

Since there are no data about the protective role of selenium (Se) against cadmium (Cd)-induced oxidative damage in early life, we studied the effect of Se supplementation on antioxidative enzyme activity and lipid peroxidation (through thiobarbituric acid reactive substances; TBARS) in suckling Wistar rats exposed to Cd. Treated animals received either Se alone for 9 days (8 μmol, i.e.

Phosphorothioate-based ligase-independent gene cloning (PLICing): An enzyme-free and sequence-independent cloning method.

Many ligase-independent cloning methods have been developed to overcome problems of standard restriction cloning such as low transformation efficiency and high background of vector with no insert. Most of these methods are still enzyme based, require time-consuming incubation and multiple purification steps, and/or might have a low robustness in handling. Thus, with the aim to establish a robust enzyme/ligase-free method, we developed the phosphorothioate-based ligase-independent gene cloning (PLICing) method, which is based on a chemical cleavage reaction of phosphorothioate bonds in an iodine/ethanol solution.

Insights into proteolytic processing of the major peanut allergen Ara h 2 by endogenous peanut proteases.

Background: The major peanut allergens are Ara h 1, Ara h 2 and Ara h 6. Proteolytic processing has been shown to be required for the maturation process of Ara h 6. The aim of this study was to examine whether Ara h 2 undergoes proteolytic processing and, if so, whether proteolytic processing influences its ability to bind human immunoglobulin E (IgE).

The effect of dietary selenium supplementation on cadmium absorption and retention in suckling rats.

Selenium (Se) reduces cadmium (Cd) toxicity in adult animals, but its effects in newborn animals are still unknown. This study investigated Cd (as CdCl2) absorption, distribution, and retention in suckling rats receiving oral Se supplementation (as Na2SeO3) in equimolar doses (8 μmol Cd and/or Se per kg b.w.

Influence of selenium dose on mercury distribution and retention in suckling rats.

It is well known that metal-metal interactions in the body are age-dependent. We studied the influence of increasing selenium (Se) doses on mercury (Hg) distribution and retention in the postnatal period in Hg-exposed suckling rats. Seven-day-old Wistar pups were pretreated with three different oral doses of Se as sodium selenite (6.

Comparison of calcium, magnesium, sodium, potassium, zinc, and creatinine concentration in 24-h and spot urine samples in women.

Background: The 24-h urine sample is considered as the most reliable material for testing many but not necessarily all constituents in urine. However, its collection is tedious for both patients and research participants. The aim of this study was to compare concentrations of essential elements calcium (Ca), magnesium (Mg), sodium (Na), potassium (K), and zinc (Zn) in 24-h and spot urine samples.

Nutrition and lifestyle in relation to bone health and body weight in Croatian postmenopausal women.

The objective was to investigate the association of nutrients and lifestyle modifiers with bone mineral density (BMD) and weight and/or body mass index (BMI) in 120 healthy Croatian postmenopausal women. The hip and spine BMD was assessed by Lunar Prodigy (GE Medical Systems). Nutrient assessment from 3-day records was analyzed using the US Department of Agriculture Food Composition Tables and the Croatian National Institute of Public Health database.

Peak bone density in Croatian women: variations at different skeletal sites.

It is known that different skeletal sites have different peak bone mass at different times and lose bone at different rates. The purpose of the study was to assess bone mineral density (BMD) in healthy female student population (N=220), aged 18-25 yr and to analyze whether young women of that age have already started to lose the bone mass at the trabecular and cortical parts of skeleton. The influence of dietary intake and physical activity on their bone mass was also assessed.

Laboratory evolution of P450 BM3 for mediated electron transfer yielding an activity-improved and reductase-independent variant.

One of the main obstacles in employing P450 monooxygenases for preparative chemical syntheses in cell-free systems is their requirement for cofactors such as NAD(P)H. In order to engineer P450 BM3 from Bacillus megaterium for cost-effective process conditions in vitro, a validated medium throughput screening system based on cheap Zn dust as an electron source and Cobalt(III)sepulchrate (Co(III)sep) as a mediator was reported. In the current study, the alternative cofactor system Zn/Co(III)sep was used in a directed evolution experiment to improve the Co(III)sep-mediated electron transfer to P450 BM3.

Toxic and essential metal concentrations in four tissues of red deer (Cervus elaphus) from Baranja, Croatia.

Kidney (n = 297), liver (n = 52), jawbone (n = 80) and muscle (n = 48) samples collected from red deer (Cervus elaphus) from north-eastern Croatia in the 2002--05 hunting season were analysed for cadmium (Cd), copper (Cu), iron (Fe), mercury (Hg), lead (Pb), selenium (Se) and zinc (Zn) using atomic absorption spectrometry. Statistical evaluation of results showed age-related accumulations of renal cortex Cd and Zn, bone Pb, and muscle Zn. Renal cortex Cd and Zn were significantly associated.

Quantification of Art v 1 and Act c 1 being major allergens of mugwort pollen and kiwi fruit extracts in mass-units by ion-exchange HPLC-UV method.

A simple ion-exchange HPLC-UV method was developed for determination of major allergens from mugwort pollen and kiwi fruit extracts in mass-units. The separation of Art v 1 and Act c 1 from other components in the extracts was achieved in one step. The extinction coefficients used in the study were theoretically determined and compared to the extinction coefficients determined by gravimetry.

Biological availability of selenosugars in rats.

The biological availability and metabolism of two selenosugars orally administered to rats were investigated. Two other selenium species, selenite and trimethylselenonium ion (TMSe) were included in the study as positive and negative controls, respectively. Male Wistar strain rats (three per group) at 8 weeks of age were exposed to sodium selenite, TMSe, selenosugar 1 (methyl-2-acetamido-2-deoxy-1-seleno-beta-D-galactopyranoside) or selenosugar 2 (methyl-2-acetamido-2-deoxy-1-seleno-beta-D-glucopyranoside) through drinking water for 48 h.

A matrix effect in pectin-rich fruits hampers digestion of allergen by pepsin in vivo and in vitro.

Background: It is a general belief that a food allergen should be stable to gastric digestion. Various acidic plant polysaccharides, including pectin, are ubiquitous in fruit matrixes and can form hydrogels under low-pH conditions.

Objective: The purpose of this study was to investigate the effect of hydrogel forming polysaccharide-rich fruit matrixes on in vivo gastric and in vitro pepsic digestion of fruit allergens.

Comparison of organic and inorganic mercury distribution in suckling rat.

Thiomersal is used as a preservative in vaccines given to small children. The metabolic product of thiomersal is ethylmercury and its distribution and kinetics are still not known, especially at this early age. The purpose of this study was to compare the body distribution of two forms of mercury: organic (thiomersal) and inorganic (mercury(2+) chloride) in very young, suckling rats.

Heavy metal concentrations in ground beetles, leaf litter, and soil of a forest ecosystem.

The objective of this study was to quantify the relationships between heavy metal concentrations in soil, leaf litter, and ground beetles at four sampling sites of a forest ecosystem in Medvednica Nature Park, Croatia. Ground beetles were sampled by pitfall trapping. Specimens were dry-ashed and soil and beetle samples digested with nitric acid.

Chelators as antidotes of metal toxicity: therapeutic and experimental aspects.

The effects of chelating drugs used clinically as antidotes to metal toxicity are reviewed. Human exposure to a number of metals such as lead, cadmium, mercury, manganese, aluminum, iron, copper, thallium, arsenic, chromium, nickel and platinum may lead to toxic effects, which are different for each metal. Similarly the pharmacokinetic data, clinical use and adverse effects of most of the chelating drugs used in human metal poisoning are also different for each chelating drug.

Mercury chloride genotoxicity in rats following oral exposure, evaluated by comet assay and micronucleus test.

Mercury is a toxic element which is easily absorbed after ingestion or inhalation and deposited mainly in the kidney. The aim of this study was to evaluate the effects of mercury chloride in rats. Female rats, aged 14 weeks, were receiving mercury chloride in oral doses of 0.

Sodium and calcium intakes and bone mass in rats revisited.

Objective: High sodium intake accompanied by insufficient dietary calcium may have detrimental effects on bone mass. Our study evaluated the effects of increased sodium and decreased calcium intakes on bone mineral density (BMD) and bone mineral content (BMC) in rats.

Methods: Four-month-old female Wistar rats were given deionized water or 1.