Preparation of monoclonal antibody bank against whole water-soluble proteins from rapid-growing bamboo shoots.

An antibody bank against the whole proteins in a proteome is a useful tool for biological research. Using the standard cell fusion method, and a modified screening protocol, we produced an mAb bank against the total water-soluble proteins extracted from the rapid-growing green bamboo shoots. An improved two-stage strategy was employed to enrich those poor immunogenic or lower expressed proteins. Totally, we obtained a bank of 192 mAb which were identified as distinctive to each other by 2-DE and immunostaining.

[Glycosylation profile of selected acute phase proteins in children with chronic tonsillitis and allergic symptoms]. / Glikozylacja bialek ostrej fazy u dzieci z zapaleniem migdalków podniebiennych i objawami alergii.

INTRODUCTION: Acute phase proteins may be regarded as laboratory markers of inflammatory processes of various origin, but they also play several important biological roles. As majority of them are glycoproteins alterations in glycosylations profiles form additional sign of disturbances in the cytokines network during inflammation and allow to distinguish between acute and chronic inflammatory conditions. MATERIAL AND METHODS: A group of 25 children, aged from 6 to 13 years, admitted due to tonsillectomy was examined using skin tests towards specific allergens. Fifteen children out of the whole group showed reaction to pollens, whereas in ten children no allergen was detected despite clear allergic symptoms. In sera samples from every child concentrations of C-reactive protein, alpha1-acid glycoprotein (AGP) and alpha1-antichymotrypsin (ACT) were measured using rocket immunoelectrophoresis acc. to Laurell, and glycosylations profiles of AGP and ACT were determined, using crossed affino-immunoelectrophoresis acc. to Bøg-Hansen. RESULTS: Lower concentration of AGP and higher of ACT was shown for children allergic to pollens. Glycosylation profile of both proteins was altered towards higher reactivity with ConA for children allergic to pollens, whereas rather chronic image was observed in children allergic to unknown allergen. The latter image was similar to previously described in children with food allergies. CONCLUSIONS: The presence of allergic reaction may alter the cytokine network activity in children, thus affecting also the immune status, independently from chronic inflammatory process in tonsillitis.

Antibody response to Pseudomonas aeruginosa in cystic fibrosis patients: a marker of therapeutic success?--A 30-year cohort study of survival in Danish CF patients after onset of chronic P. aeruginosa lung infection.

We studied the effects of increasingly intensive treatment regimens on anti-pseudomonal antibody response and survival in five successive cohorts of a total of 157 Danish cystic fibrosis patients after they had acquired chronic P. aeruginosa lung infection. The time periods were 1971-1975 (N = 21), 1976-1980 (N = 64), 1981-1986 (N = 27), 1987-1993 (N = 26), and 1994-2000 (N = 19). During this 30-year period, we introduced elective 2-week courses of chemotherapy every third month in all chronically infected patients, early aggressive treatment with inhalation of colistin and oral ciprofloxacin for 3 months whenever P. aeruginosa was cultured in sputum from noncolonized patients, and inhalation of recombinant human dornase alfa. There was a significant correlation between the calendar year when chronic P. aeruginosa infection was acquired and the subsequent increase in the level of precipitins (P < 0.00001). The median number of precipitins increased by 5 per year in the oldest calendar year cohort, and 1 per year in the youngest. The median age of onset of chronic P. aeruginosa increased from 9.3 years from 1981-1986 to 13.8 years from 1987-2000. Survival after acquisition of chronic P. aeruginosa lung infection improved with time (P = 0.008). Our study shows that CF patients who are treated intensively have lower antibody responses and longer survival after acquisition of chronic P. aeruginosa lung infection.

Standardisation of Glutaraldehyde-modified Tyrosine-adsorbed Tree Pollen Vaccines Containing the Th1-inducing Adjuvant, Monophosphoryl Lipid A (MPL)

Background: a new range of allergy vaccines has been developed by the introduction of a relatively new Th1-inducing adjuvant known as 3-deacylated monophosphoryl lipid A (MPL®). MPL® adjuvant is of natural origin, derived from the lipopolysaccharide of Salmonella minnesota R595. This adjuvant is incorporated in a glutaraldehyde-modified pollen extract adsorbed to L-tyrosine (Pollinex® Quattro). A major potential benefit provided by MPL® adjuvant is the promotion of a Th1 response which enhances the efficacy of allergy vaccination and can consequently allow a reduction in the number of injections required for treatment. The standardisation of Pollinex® Quattro tree pollen allergy vaccine is described and we include details of some innovative analytical procedures. Methods and results: an essential feature of the analytical strategy is the assay of the MPL® adjuvant using a recently developed HPLC technique. The adjuvant has a complex chemical structure and the analysis is illustrated in detail. We give a full picture of the vaccine standardisation by describing biochemical and immunological characterisation of the allergen extract, together with some brief manufacturing details. Conclusions: a high overall level of standardisation is illustrated by a number of different tests applied to all stages of vaccine manufacture. Tree pollen allergen potency is measured following the pollen extraction, chemical modification and formulation as a tyrosine adsorbate. Good batch-to-batch reproducibility is shown. The HPLC assay for MPL® adjuvant showed high quality resolution which did not vary when measuring raw material or when incorporated in the vaccine and the technically complex assay is shown to be reliable (AU)

Antecedentes: un nuevo tipo de Inmunoterapia específica (I.T.s.) ha sido desarrollado por la introducción de un nuevo adyuvante, MPL (3-deacylated monophosphoryl lipid A), inductor Th1.MPL es un adyuvante de origen natural, derivado del lipopolisacárido de la Salmonela minnesota R595.Este adyuvante se incorpora a un extracto de pólenes modificados con glutaraldehído y adsorbidos con tirosina (Pollinex Quattro).El principal beneficio potencial proporcionado por el adyuvante MPL consiste en la inducción de la respuesta Th1 que incrementa la eficacia de la I.T.s y, consecuentemente, permite una reducción en el número de inyecciones requerido para el tratamiento. Se describe la estandarización de Pollinex Quattro con pólenes de árboles y se incluyen detalles de algunos procedimientos analíticos innovativos. Métodos y resultados: una característica esencial de la estrategia analítica consiste en la valoración del MPL utilizando una técnica HPLC recientemente desarrollada. El adyuvante tiene una estructura química compleja cuyo análisis se describe en detalle. Se proporciona una completa descripción de la estandarización de la vacuna basada en la caracterización bioquímica e inmunológica del extracto alergénico, junto a unos detalles técnicos de elaboración. Conclusiones: la alta calidad de la metodología de estandarización se ve reflejada por los diferentes tests aplicados a todas las fases del proceso de elaboración de la vacuna. La potencia alergénica de los pólenes de árboles se determina en cada una de las fases: extracción del polen, modificación química y formulación con tirosina como adsorbente. Se demuestra la buena reproductibilidad lote a lote. El ensayo HPLC para MPL presentó una resolución de alta calidad que no variaba cuando se determinaba como materia prima o una vez incorporado a la vacuna; mostrándose la compleja valoración técnica fiable (AU)

Standardisation of glutaraldehyde-modified tyrosine-adsorbed tree pollen vaccines containing the Th1-inducing adjuvant, monophosphoryl lipid A (MPL).

BACKGROUND: a new range of allergy vaccines has been developed by the introduction of a relatively new Th1-inducing adjuvant known as 3-deacylated monophosphoryl lipid A (MPL). MPL adjuvant is of natural origin, derived from the lipopolysaccharide of Salmonella minnesota R595. This adjuvant is incorporated in a glutaraldehyde-modified pollen extract adsorbed to L-tyrosine (Pollinex Quattro). A major potential benefit provided by MPL adjuvant is the promotion of a Th1 response which enhances the efficacy of allergy vaccination and can consequently allow a reduction in the number of injections required for treatment. The standardisation of Pollinex Quattro tree pollen allergy vaccine is described and we include details of some innovative analytical procedures. METHODS AND RESULTS: an essential feature of the analytical strategy is the assay of the MPL adjuvant using a recently developed HPLC technique. The adjuvant has a complex chemical structure and the analysis is illustrated in detail. We give a full picture of the vaccine standardisation by describing biochemical and immunological characterisation of the allergen extract, together with some brief manufacturing details. CONCLUSIONS: a high overall level of standardisation is illustrated by a number of different tests applied to all stages of vaccine manufacture. Tree pollen allergen potency is measured following the pollen extraction, chemical modification and formulation as a tyrosine adsorbate. Good batch-to-batch reproducibility is shown. The HPLC assay for MPL adjuvant showed high quality resolution which did not vary when measuring raw material or when incorporated in the vaccine and the technically complex assay is shown to be reliable.

Alpha-1 antichymotrypsin levels are actively increased in normal colostrum.

BACKGROUND: It is well known that human colostrum has important antiinflammatory functions. The purpose of the current study was to determine antiprotease levels in colostrum and serum and to assess the importance of local synthesis and the electrophoretic differences in both locations. METHODS: Five protease inhibitors were determined by radical immunodiffusion in colostrum and serum samples taken simultaneously from 50 healthy women, 36 to 72 hours after delivery. RESULTS: Antithrombin II, inter-alpha trypsin inhibitor, and alpha-2 macroglobulin levels were undetectable in colostrum. Mean antitrypsin levels in colostrum were 6% of serum levels, but colostrum alpha-1 antichymotrypsin was higher than expected (0.39+/-0.34 g/l) in relation according to the albumin passive transport, and their mean value was 41% of serum levels. Colostrum antichymotrypsin levels did not correlate with serum antichymotrypsin levels or with colostrum albumin levels. The antichymotrypsin molecule in colostrum had a slower electrophoretic mobility when compared with that of serum antichymotrypsin, and it showed a different pattern in Western blot analysis, with a predominating 80-kDa molecule. CONCLUSIONS: Although the origin of colostral antichymotrypsin is unclear, local production in breast epithelial cells is likely. Antichymotrypsin is increased in colostrum, and its molecule has some characteristic differences, suggesting that it has an important and specific role in infant nutrition during breast milk feeding.

Comparison of allergenic potentials of timothy (Phleum pratense) pollens from different pollen seasons collected in the Belgrade area.

We investigated extracts of timothy grass pollen from four seasons (1989, 1990, 1991, and 1994) by protein content, SDS-PAGE, immunoblot, RAST, RAST inhibition, and crossed immunoelectrophoresis. Extract of the pollen from 1991 showed the lowest yield in quantitative assays. SDS-PAGE, crossed immunoelectrophoresis, RAST, and RAST inhibition expressed approximately comparable patterns for all extracts except that from 1991. Obviously, the quality of grass pollens, as shown for some ragweed (Ambrosia elatior) pollens depend on year of collection. Our findings are partially in agreement with some earlier examinations of the quality of timothy pollen from different pollen seasons.

Comparison between outdoor and indoor airborne allergenic activity.

BACKGROUND: Allergenic pollens are usually detected in outdoor air by using volumetric spore traps, which allow measurement of atmospheric concentration as pollen grains per m3 of air. The results of the pollen count are useful primarily for outdoor environments while most people spend most of the day indoors. OBJECTIVE: The purpose of our study was to compare outdoor pollen levels with allergenic activity measured both outdoors and indoors. METHODS: We used a Lanzoni spore trap to measure airborne Urticaceae pollen and filters collecting particles simultaneously indoors and outdoors and assayed each filter for Parietaria judaica allergenic activity. Samples were collected at the Allergological Service of the "A. Cardarelli" Hospital in Naples with the balcony open on some days and closed on others. Allergenic activity (ng/m3) was measured using the immunocapture RAST. RESULTS: With the balcony open there was no great difference between outdoor and indoor allergenic activity, but with the balcony closed there was a reduction of indoor allergenic activity of about one-third in comparison with outdoor allergenic activity. Statistical analysis (Pearson correlation test) indicated a significant correlation between outdoor allergen levels and indoor allergen levels with the balcony open (r = .4415, P < .05), but not with the balcony closed (r = .3160, P > .05); a significant correlation between outdoor pollen count and indoor allergen levels with the balcony open (r = .4809, P < .05), but not with the balcony closed (r = .3858, P > .05); and a highly significant correlation (r = .5225, P < .001) between outdoor pollen count and outdoor allergen levels. CONCLUSIONS: These data provide scientific evidence for the recommendation to hay fever patients to remain indoors during seasons with high levels of outdoor pollens.

Interactions between phytoestrogens and human sex steroid binding protein.

The interactions of human Sex steroid binding protein (SBP) and the lignans [Nordihydrogaiaretic acid (NDGA) enterolactone (Ent), enterodiol (End)] and isoflavonoid phytoestrogens [Equol (Eq), diazein Dad), genistein (Gen)] were studied. The phytoestrogens had different dose-dependent inhibitory effects on steroid binding by SBP. Their relative efficiencies were: Ent> or = NDGA = Eq > Gen for displacing E2 and Eq > Ent > NDGA > Gen for displacing T. End and Dad were much less active. Scatchard analysis suggested that NDGA had similar non- competitive effects on T and E2 binding by reducing the number of binding sites without changing the association constants. But Eq seemed to inhibit E2 binding non-competitively and T binding competitively. NDGA binding to SBP reduced the immunorecognition of SBP by monospecific anti-SBP antibodies, suggesting that NDGA changed SBP immunoreactivity. Unlike NDGA, Eq binding to SBP caused no immunological changes in SBP, indicating qualitative differences in the effects of the lignan and isoflavonoid. Our results indicate that phytoestrogens may modulate the SBP activity and so influence the role of this protein in the delivery of hormonal information to sex steroid-dependent cells.

English plantain and psyllium: lack of cross-allergenicity by crossed immunoelectrophoresis.

BACKGROUND: English plantain (Plantago lanceolata) weed pollen and psyllium (Plantago ovata) husk dust are inhalant allergens. Because of the phylogenetic relationship between these plant species, cross-allergenicity has been a concern. OBJECTIVE: The purpose of this study was to investigate the possible cross-allergenicity of plantain and psyllium. METHODS: Homologous and heterologous crossed immunoelectrophoresis (CIE) were performed using a commercial English plantain pollen extract and an extract of psyllium seed embryo. Crossed radioimmunoelectrophoresis (CRIE) was performed using sera from subjects who were RAST positive only to plantain (group A), RAST positive only to psyllium (group B), RAST positive to both plantain and psyllium (group C), or RAST negative to both (group D). RESULTS: All of the group A plantain subjects showed IgE binding to at least one of the six plantain allergens in homologous plantain CRIEs while only one of the sera from the group B subjects reacted very weakly to these plantain allergens. In homologous psyllium CRIE, all group B subjects showed pronounced IgE binding to 2 to 7 of the seven psyllium allergens. Several of the plantain subjects demonstrated only very weak binding to psyllium allergens. Heterologous CRIEs demonstrated little relevant IgE binding. CONCLUSIONS: These results indicate that there is little cross-allergenicity between psyllium husk and English plantain pollen.

[The isolation and purification of the protective antigen and the edema factor from the culture filtrate of Bacillus anthracis STI-1]. / Vydelenie i ochistka protektivnogo antigena i otechnogo faktora iz kul'tural'nogo fil'trata Bacillus anthracis STI-1.

A rapid method for producing a protective antigen (PA) and edema factor (EF), components of anthrax toxin, is described. The specific features of the method are as follows: addition of a mixture of protease inhibitors to the culture fluid; simultaneous concentration on a fiber filter and adsorption on hydroxylapatite, followed by non-linear gradient a phosphate concentration; purification of elution of a phosphate concentration; purification of eluates from salts via electrodialysis. Resultant from protein desorption, PA and EF agents are electrophoretically homogeneous by no less than 80%, 10 liters of filtrate yielded 8 mg of PA and 5 mg of EF. The resultant agents were tested for their biological properties and protective activity.

Characterization of Chenopodiales (Amaranthus retroflexus, Chenopodium album, Kochia scoparia, Salsola pestifer) pollen allergens.

Pollen extracts of the four taxonomically related weeds, Amaranthus retroflexus (Ama r), Chenopodium album (Che a), Kochia scoparia (Koc s), and Salsola pestifer (S. kali) (Sal p), were characterized by various methods including crossed immunoelectrophoresis (CIE), crossed radioimmunoelectrophoresis (CRIE), and SDS-PAGE immunoblotting. The allergen profiles were determined by CRIE and SDS-PAGE IgE immunoblotting. CRIE detected from one to four important allergens, while SDS showed up to four bands that bound IgE from a number of patient sera. CRIE and SDS-PAGE immunoblotting did not recognize the same number of important allergens in the individual weeds, and the number of allergens detected by the two methods differed considerably, suggesting that IgE-binding epitopes may be denatured during SDS-PAGE. However, it was possible to correlate the identity of some of the important allergens detected by CRIE and SDS-PAGE immunoblotting in all four weeds.

Antigenic relationship between mugwort and ragweed pollens by crossed immunoelectrophoresis.

Mugwort and ragweed pollens have been considered as important respiratory allergens in Korea. These two pollens are abundant in the air of Seoul from August through October. Many ragweed-sensitive patients have shown concurrent sensitivities to mugwort pollen. However the antigenic relationship between these two pollens has not been clarified. To observe the cross-reactivity between them, we developed polyclonal anti-mugwort and anti-ragweed antibodies by immunization on New Zealand white rabbits, and performed crossed immunoelectrophoresis(CIE) with two pollen extracts. Five precipitation lines were formed by mugwort and anti-mugwort antibody. One precipitation line was formed by ragweed and anti-ragweed antibody. There was no reaction from mugwort and anti-ragweed antibody, and from ragweed and anti-mugwort antibody. These results indicate that there is no cross-antigenicity between mugwort and ragweed pollens.

Antigenic relationship between mugwort and ragweed pollens by crossed immunoelectrophoresis

Mugwort and ragweed pollens have been considered as important respiratory allergens in Korea. These two pollens are abundant in the air of Seoul from August through October. Many ragweed-sensitive patients have shown concurrent sensitivities to mugwort pollen. However the antigenic relationship between these two pollens has not been clarified. To observe the cross-reactivity between them, we developed polyclonal anti-mugwort and anti-ragweed antibodies by immunization on New Zealand white rabbits, and performed crossed immunoelectrophoresis(CIE) with two pollen extracts. Five precipitation lines were formed by mugwort and anti-mugwort antibody. One precipitation line was formed by ragweed and anti-ragweed antibody. There was no reaction from mugwort and anti-ragweed antibody, and from ragweed and anti-mugwort antibody. These results indicate that there is no cross-antigenicity between mugwort and ragweed pollens.

Characterization of recombinant bet vI, the major pollen allergen of Betula verrucosa (white birch), produced by fed-batch fermentation.

The gene encoding the major allergen, Bet v I, from Betula verrucosa (white birch) pollen was cloned by application of the polymerase chain reaction with double-stranded cDNA as template and specific primers based on the published nucleotide sequence of the gene. The gene was inserted into plasmid pKK223-3 and expressed in Escherichia coli K-12 strain JM105 grown to high cell density in a fermenter using a fed-batch procedure. Ample material was provided for a thorough characterization of the epitope structure of recombinant Bet v I contained in an unpurified soluble lysate. The antibody-binding characteristics of recombinant Bet v I was compared with that of the natural allergen using polyclonal rabbit antibodies raised against Bet v I in crossed immunoelectrophoresis, tandem crossed immunoelectrophoresis, and Western blotting. IgE from a pool of 16 allergic patients' serum was applied in crossed radioimmunoelectrophoresis, Western blotting, and a quantitative luminescence inhibition immunoassay. Well-defined epitopes were assayed by the application of a panel of six murine monoclonal antibodies in a quantitative radio inhibition immunoassay. In all assays the activity of recombinant Bet v I was comparable to that of natural Bet v I, and it is concluded that the epitope structure of recombinant Bet v I closely resembles that of natural Bet v I. This result has important implications for the future use of recombinant tree pollen allergens as a model system for the study of allergenic B and T cell epitopes aiming at improvements in reagents used for the management of allergic disease.

Structural and functional microheterogeneity of rat thyroxine-binding globulin during ontogenesis.

Thyroxine-binding globulin (TBG), the major carrier of thyroid hormones in human and murine sera, is in the rat a developmentally regulated protein, showing a large surge during post-natal growth followed by virtual disappearance in adults. Here we study as a function of age, from the 19-day embryo to 60 days after birth, the structural and binding characteristics of rat TBG microheterogeneity. Serum obtained throughout development, when pre-incubated with 125I-thyroxine (T4), was shown by isoelectric focusing (IEF; pH range 4-5) to contain six labelled isoforms of TBG, with isoelectric points between 4.25 and 4.55. These isoforms differ in their sialic acid content. The relative labelling densities of the isoforms show age-related changes: in neonates, the bulk of T4 is bound to the most alkaline (least sialylated) TBG isoforms; then, with advancing age, it shifts to the most acidic isoforms. To understand whether this progressive transfer of ligand reflects developmental changes in the relative abundance of isoforms, we submitted sera from rats of different ages to crossed immunoelectrofocusing analysis. We demonstrate that the relative proportions of the TBG isoforms remain fairly constant, independent of the level of total TBG. The most acidic forms always represented the majority (approximately 50%), with the most alkaline ones only representing 15% of total TBG. Experiments based on IEF of charcoal-treated sera, supplemented or not with lipidic serum extracts, further demonstrate that the paradoxical low labelling seen in the neonates for the most abundant highly sialylated isoforms is due to inhibition of their binding abilities by liposoluble components, which are particularly concentrated in the sera at the earlier post-natal ages. These studies represent the first analysis of concentration versus binding functions of rat TBG isoforms in the physiological conditions of normal ontogeny. Our results point to an important influence for the serum environment on the binding properties of TBG isoforms. The physiological significance of such interactions remains to be clarified.

PCR based cloning and sequencing of isogenes encoding the tree pollen major allergen Car b I from Carpinus betulus, hornbeam.

Cloning of the gene encoding the major allergen, Car b I, from Carpinus betulus (hornbeam) pollen was performed using the Polymerase Chain Reaction (PCR) to specifically amplify the gene of interest using single stranded cDNA as template. Specific primers, deduced from the aminoterminal sequence of the purified protein, were tailored to facilitate direct expression of plasmic clones, and the large fraction of positive clones obtained, revealed the presence of isogenic variation. Three clones were characterized in detail by antibody based assays and nucleotide sequencing. The recombinant allergens were shown by crossed immunoelectrophoresis (CIE) to precipitate with monospecific polyclonal rabbit antibodies raised against purified Bet v I, by crossed radioimmunoelectrophoresis (CRIE) to bind tree pollen allergic patient serum IgE, and by immunoblotting to bind murine monoclonal antibodies, raised against purified Car b I from pollen. Car b I is encoded by a 159-triplets open reading frame. The molecular masses (M(r) = 17272, 17355 and 17217 Da, respectively), the amino acid composition, and the aminoterminal sequence of the predicted polypeptides agree well with data obtained by analysis of the protein purified from pollen. The deduced amino acid sequences show pronounced homology (73, 75 and 74% identities respectively) to Bet v I, the major allergen from Betula verrucosa (white birch) pollen. Soluble recombinant Car b I, without a fusion partner, was produced in Escherichia coli with an immunochemical reactivity closely resembling that of the native pollen allergen. The tree pollen major allergens therefore constitute an ideal system for the study of allergenic epitopes.

Antigenic and allergenic analysis of psyllium seed components.

The outer portions (husk) of psyllium seeds are a concentrated source of natural fiber used in some bulk-fiber laxatives and cereals. They are known to elicit respiratory allergic reactions after inhalation or ingestion among sensitized individuals. Antigenic and allergenic characterization of three psyllium-seed fractions (husk, endosperm, and embryo) was conducted with crossed immunoelectrophoresis (CIE), crossed radioimmunoelectrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine the source of psyllium allergenicity. Homologous CIE demonstrated psyllium endosperm and embryo extracts contained seven and four antigens, respectively. Husk extracts were too gelatinous to react by CIE. However, heterologous CIE profiles of endosperm or embryo extracts, reacted with antihusk antibodies, resulted in antigen-antibody precipitin peaks that matched the heavy staining precipitin lines of homologous reactions for endosperm and embryo, respectively. These results indicated that commercial-grade husk, endosperm, and embryo contained similar antigens. Extracts of all three seed components contained antigens that bound IgE antibodies in the sera of 11 psyllium RAST-positive individuals, as determined by crossed radioimmunoelectrophoresis. The few prominent husk protein/peptide bands resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were common in either embryo or endosperm. Immunoblots revealed common IgE reactive bands in all three seed fractions. Microscopic examination of the powdered commercial-grade psyllium (95% pure) revealed it contained endosperm and embryo particles. These immunologic, biochemical, and microscopic findings suggest that other contaminating seed components are primarily responsible for the allergenicity of commercial-grade psyllium-husk powder rather than the husk itself.

[A comparative immunochemical analysis of allergoids and allergens]. / Sravnitel'nyi immunokhimicheskii analiz allergoidov i allergenov.

In comparison with allergens having protein fragments with a molecular weight not exceeding 110 kD, allergoids have been found to consist of larger fragments with a molecular weight of 10-150 kD. Allergoids have less charged components than initial allergens and less antigenic components. Allergoids retain their capacity for stimulating the production of antibodies, specific to all antigenic components.

Purification of Phleum pratense pollen extract by immunoaffinity chromatography and high-performance ion-exchange chromatography.

Basic allergens of Phleum pratense pollen extract have been purified by either sequence gel filtration-ion-exchange high-performance liquid chromatography (HPIEX) and size-exclusion HPLC or sequence gel filtration-immunoaffinity chromatography and HPIEX. The second procedure seems to be suitable for preparative purposes.