TY  - JOUR
A1  - Roggenbuck, Dirk
A1  - Borghi, Maria Orietta
A1  - Somma, Valentina
A1  - Buettner, Thomas
A1  - Schierack, Peter
A1  - Hanack, Katja
A1  - Grossi, Claudia
A1  - Bodio, Caterina
A1  - Macor, Paolo
A1  - von Landenberg, Philipp
A1  - Boccellato, Francesco
A1  - Mahler, Michael
A1  - Meroni, Pier Luigi
T1  - Antiphospholipid antibodies detected by line immunoassay differentiate among patients with antiphospholipid syndrome, with infections and asymptomatic carriers
JF  - IEEE transactions on geoscience and remote sensing
N2  - Background: Antiphospholipid antibodies (aPL) can be detected in asymptomatic carriers and infectious patients. The aim was to investigate whether a novel line immunoassay (LIA) differentiates between antiphospholipid syndrome (APS) and asymptomatic aPL+ carriers or patients with infectious diseases (infectious diseases controls (IDC)). Methods: Sixty-one patients with APS (56 primary, 22/56 with obstetric events only, and 5 secondary), 146 controls including 24 aPL+ asymptomatic carriers and 73 IDC were tested on a novel hydrophobic solid phase coated with cardiolipin (CL), phosphatic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, beta2-glycoprotein I (beta 2GPI), prothrombin, and annexin V. Samples were also tested by anti-CL and anti-beta 2GPI ELISAs and for lupus anticoagulant activity. Human monoclonal antibodies (humoAbs) against human beta 2GPI or PL alone were tested on the same LIA substrates in the absence or presence of human serum, purified human beta 2GPI or after CL-micelle absorption. Results: Comparison of LIA with the aPL-classification assays revealed good agreement for IgG/IgM a beta 2GPI and aCL. Anti-CL and anti-beta 2GPI IgG/IgM reactivity assessed by LIA was significantly higher in patients with APS versus healthy controls and IDCs, as detected by ELISA. IgG binding to CL and beta 2GPI in the LIA was significantly lower in aPL+ carriers and Venereal Disease Research Laboratory test (VDRL) + samples than in patients with APS. HumoAb against domain 1 recognized beta 2GPI bound to the LIA-matrix and in anionic phospholipid (PL) complexes. Absorption with CL micelles abolished the reactivity of a PL-specific humoAb but did not affect the binding of anti-beta 2GPI humoAbs. Conclusions: The LIA and ELISA have good agreement in detecting aPL in APS, but the LIA differentiates patients with APS from infectious patients and asymptomatic carriers, likely through the exposure of domain 1.
KW  - Antiphospholipid syndrome
KW  - Antiphospholipid antibody
KW  - Phospholipid binding proteins
KW  - Beta2-glycoprotein I
KW  - Line immunoassay
Y1  - 2016
U6  - https://doi.org/10.1186/s13075-016-1018-x
SN  - 1478-6354
SN  - 1478-6362
VL  - 18
PB  - BioMed Central
CY  - London
ER  - 
TY  - GEN
A1  - Hanack, Katja
A1  - Schloer, Anja
A1  - Holzloehner, Pamela
A1  - Listek, Martin
A1  - Bauer, Cindy
A1  - Butze, Monique
A1  - Micheel, Burkhard
A1  - Hentschel, Christian
A1  - Sowa, Mandy
A1  - Roggenbuck, Dirk
A1  - Schierack, Peter
A1  - Fuener, Jonas
A1  - Schliebs, Erik
A1  - Goihl, Alexander
A1  - Reinhold, Dirk
T1  - Camelid nanobodies specific to human pancreatic glycoprotein 2
T2  - The journal of immunology
N2  - Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified to be a major autoantigenic target in Crohn’s disease patients. It was discussed recently that a long and a short isoform of GP2 exists whereas the short isoform is often detected by GP2-specific autoantibodies. In the outcome of inflammatory bowel diseases, these GP2-specific autoantibodies are discussed as new serological markers for diagnosis and therapeutic monitoring. To investigate this further, camelid nanobodies were generated by phage display and selected against the short isoform of GP2 in order to isolate specific tools for the discrimination of both isoforms. Nanobodies are single domain antibodies derived from camelid heavy chain only antibodies and characterized by a high stability and solubility. The selected candidates were expressed, purified and validated regarding their binding properties in different enzyme-linked immunosorbent assays formats, immunofluorescence, immunohistochemistry and surface plasmon resonance spectroscopy. Four different nanobodies could be selected whereof three recognize the short isoform of GP2 very specifically and one nanobody showed a high binding capacity for both isoforms. The KD values measured for all nanobodies were between 1.3 nM and 2.3 pM indicating highly specific binders suitable for the application as diagnostic tool in inflammatory bowel disease.
Y1  - 2016
SN  - 0022-1767
SN  - 1550-6606
VL  - 196
SP  - 313
EP  - 328
PB  - American Assoc. of Immunologists
CY  - Bethesda
ER  - 
TY  - JOUR
A1  - Roggenbuck, Dirk
A1  - Goihl, Alexander
A1  - Hanack, Katja
A1  - Holzloehner, Pamela
A1  - Hentschel, Christian
A1  - Veiczi, Miklos
A1  - Schierack, Peter
A1  - Reinhold, Dirk
A1  - Schulz, Hans-Ulrich
T1  - Serological diagnosis and prognosis of severe acute pancreatitis by analysis of serum glycoprotein 2
JF  - Clinical chemistry and laboratory medicine : journal of the Forum of the European Societies of Clinical Chemistry - the European Branch of the International Federation of Clinical Chemistry and Laboratory Medicine
N2  - To better understand emerging adults’ perceptions of family interactions and value transmission to the next generation, we examined Hmong American emerging adults’ reflections on their parents’ parenting. Participants discussed what parenting practices they would do differently and others they hoped to emulate with their future adolescent children. Thirty Hmong American emerging adults (18-25 years; M = 21.2 years; 50% female) participated in interviews that focused retrospectively on the parent–adolescent relationship. Results revealed that emerging adults wanted to parent differently in three ways: less pressure about education, fewer restrictions, and more open communication. Emerging adults imagined being a similar
parent in four ways: promoting education, promoting life values, giving
guidance, and offering love and support. The findings highlight parenting practices that Hmong American emerging adults plan on transmitting (and not transmitting) to their own children, offering a glimpse into the type of parents the emerging adults may become.
KW  - acute pancreatitis
KW  - chronic pancreatitis
KW  - GP2 isoform alpha
KW  - pancreatic neoplasms
KW  - severe acute pancreatitis
KW  - zymogen granule membrane glycoprotein GP2
Y1  - 2017
U6  - https://doi.org/10.1515/cclm-2016-0797
SN  - 1434-6621
SN  - 1437-4331
VL  - 55
SP  - 854
EP  - 864
PB  - De Gruyter
CY  - Berlin
ER  - 
TY  - JOUR
A1  - Schiebel, Juliane
A1  - Boehm, Alexander
A1  - Nitschke, Joerg
A1  - Burdukiewicz, Michal
A1  - Weinreich, Joerg
A1  - Ali, Aamir
A1  - Roggenbuck, Dirk
A1  - Roediger, Stefan
A1  - Schierack, Peter
T1  - Genotypic and Phenotypic Characteristics Associated with Biofilm Formation by Human Clinical Escherichia coli Isolates of Different Pathotypes
JF  - Applied and environmental microbiology
N2  - Bacterial biofilm formation is a widespread phenomenon and a complex process requiring a set of genes facilitating the initial adhesion, maturation, and production of the extracellular polymeric matrix and subsequent dispersal of bacteria. Most studies on Escherichia coli biofilm formation have investigated nonpathogenic E. coli K-12 strains. Due to the extensive focus on laboratory strains in most studies, there is poor information regarding biofilm formation by pathogenic E. coli isolates. In this study, we genotypically and phenotypically characterized 187 human clinical E. coli isolates representing various pathotypes (e.g., uropathogenic, enteropathogenic, and enteroaggregative E. coli). We investigated the presence of biofilm-associated genes ("genotype") and phenotypically analyzed the isolates for motility and curli and cellulose production ("phenotype"). We developed a new screening method to examine the in vitro biofilm formation ability. In summary, we found a high prevalence of biofilm-associated genes. However, we could not detect a biofilm-associated gene or specific phenotype correlating with the biofilm formation ability. In contrast, we did identify an association of increased biofilm formation with a specific E. coli pathotype. Enteroaggregative E. coli (EAEC) was found to exhibit the highest capacity for biofilm formation. Using our image-based technology for the screening of biofilm formation, we demonstrated the characteristic biofilm formation pattern of EAEC, consisting of thick bacterial aggregates. In summary, our results highlight the fact that biofilm-promoting factors shown to be critical for biofilm formation in nonpathogenic strains do not reflect their impact in clinical isolates and that the ability of biofilm formation is a defined characteristic of EAEC. IMPORTANCE Bacterial biofilms are ubiquitous and consist of sessile bacterial cells surrounded by a self-produced extracellular polymeric matrix. They cause chronic and device-related infections due to their high resistance to antibiotics and the host immune system. In nonpathogenic Escherichia coli, cell surface components playing a pivotal role in biofilm formation are well known. In contrast, there is poor information for their role in biofilm formation of pathogenic isolates. Our study provides insights into the correlation of biofilm-associated genes or specific phenotypes with the biofilm formation ability of commensal and pathogenic E. coli. Additionally, we describe a newly developed method enabling qualitative biofilm analysis by automated image analysis, which is beneficial for high-throughput screenings. Our results help to establish a better understanding of E. coli biofilm formation.
KW  - biofilm formation
KW  - Escherichia coli
KW  - pathotypes
KW  - VideoScan
Y1  - 2017
U6  - https://doi.org/10.1128/AEM.01660-17
SN  - 0099-2240
SN  - 1098-5336
VL  - 83
PB  - American Society for Microbiology
CY  - Washington
ER  - 
TY  - JOUR
A1  - Schlör, Anja
A1  - Holzlöhner, Pamela
A1  - Listek, Martin
A1  - Grieß, Cindy
A1  - Butze, Monique
A1  - Micheel, Burkhard
A1  - Hentschel, Christian
A1  - Sowa, Mandy
A1  - Roggenbuck, Dirk
A1  - Schierack, Peter
A1  - Füner, Jonas
A1  - Schliebs, Erik
A1  - Goihl, Alexander
A1  - Reinhold, Dirk
A1  - Hanack, Katja
T1  - Generation and validation of murine monoclonal and camelid recombinant single domain antibodies specific for human pancreatic glycoprotein 2
JF  - New biotechnology
N2  - Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified as a major autoantigenic target in Crohn’s disease patients. It was reported recently that a long (GP2a) and a short (GP2b) isoform of GP2 exist and that in the outcome of inflammatory bowel diseases (IBD) GP2-specific autoantibodies probably appear as new serological markers for diagnosis and therapeutic monitoring. To investigate this further and in order to establish diagnostic tools for the discrimination of both GP2 isoforms, a set of different murine monoclonal and camelid recombinant single domain antibodies (camelid VHH) was generated and validated in various enzyme-linked immunosorbent assay (ELISA) formats, immunofluorescence on transgenic cell lines and immunohistochemistry on monkey pancreas tissue sections. Out of six binders identified, one was validated as highly specific for GP2a. This murine monoclonal antibody (mAb) was used as capture antibody in construction of a sandwich ELISA for the detection of GP2a. Camelid VHHs or a second murine mAb served as detection antibodies in this system. All antibodies were also able to stain GP2a or GP2b on transgenic cell lines as well as on pancreatic tissue in immunohistochemistry. The KD values measured for the camelid VHHs were between 7 nM and 23pM. This set of specific binders will enable the development of suitable diagnostic tools for GP2-related studies in IBD.
KW  - glycoprotein GP2
KW  - Monoclonal antibodies
KW  - Camelid single domain antibodies
Y1  - 2018
U6  - https://doi.org/10.1016/j.nbt.2018.03.006
SN  - 1871-6784
SN  - 1876-4347
VL  - 45
SP  - 60
EP  - 68
PB  - Elsevier
CY  - Amsterdam
ER  - 
TY  - JOUR
A1  - Deutschmann, Claudia
A1  - Roggenbuck, Dirk
A1  - Schierack, Peter
A1  - Rödiger, Stefan
T1  - Autoantibody testing by enzyme-linked immunosorbent assay-a case in which the solid phase decides on success and failure
JF  - Heliyon
N2  - Background: The enzyme-linked immunosorbent assay (ELISA) is an indispensable tool for clinical diagnostics to identify or differentiate diseases such as autoimmune illnesses, but also to monitor their progression or control the efficacy of drugs. One use case of ELISA is to differentiate between different states (e.g. healthy vs. diseased). Another goal is to quantitatively assess the biomarker in question, like autoantibodies. Thus, the ELISA technology is used for the discovery and verification of new autoantibodies, too. Of key interest, however, is the development of immunoassays for the sensitive and specific detection of such biomarkers at early disease stages. Therefore, users have to deal with many parameters, such as buffer systems or antigen-autoantibody interactions, to successfully establish an ELISA. Often, fine-tuning like testing of several blocking substances is performed to yield high signal-to-noise ratios. <br /> Methods: We developed an ELISA to detect IgA and IgG autoantibodies against chitinase-3-like protein 1 (CHI3L1), a newly identified autoantigen in inflammatory bowel disease (IBD), in the serum of control and disease groups (n = 23, respectively). Microwell plates with different surface modifications (PolySorp and MaxiSorp coating) were tested to detect reproducibility problems. <br /> Results: We found a significant impact of the surface properties of the microwell plates. IgA antibody reactivity was significantly lower, since it was in the range of background noise, when measured on MaxiSorp coated plates (p < 0.0001). The IgG antibody reactivity did not differ on the diverse plates, but the plate surface had a significant influence on the test result (p = 0.0005). <br /> Conclusion: With this report, we want to draw readers' attention to the properties of solid phases and their effects on the detection of autoantibodies by ELISA. We want to sensitize the reader to the fact that the choice of the wrong plate can lead to a false negative test result, which in turn has serious consequences for the discovery of autoantibodies.
KW  - biochemistry
KW  - coatings
KW  - surface chemistry
KW  - immunology
KW  - proteins
KW  - laboratory medicine
KW  - clinical research
KW  - enzyme-linked immunosorbent
KW  - assay
KW  - biomarker discovery
KW  - reproducibility
KW  - solid-phase
KW  - autoantibody
Y1  - 2020
U6  - https://doi.org/10.1016/j.heliyon.2020.e03270
SN  - 2405-8440
VL  - 6
IS  - 1
PB  - Elsevier
CY  - London [u.a.]
ER  - 
TY  - JOUR
A1  - Nawaz, Shiza
A1  - Khan, Muhammad Moman
A1  - Noack, Jonas
A1  - Awan, Asad Bashir
A1  - Schiebel, Juliane
A1  - Roggenbuck, Dirk
A1  - Schierack, Peter
A1  - Sarwar, Yasra
A1  - Ali, Aamir
T1  - Rapid detection of biofilm formation by zoonotic serovars of Salmonella enterica and avian pathogenic E. coli isolates from poultry
JF  - Pakistan veterinary journal
N2  - Biofilms are complex, sessile microbial communities that are problematic in clinical settings due to their association with survival and pathogenicity of bacteria. The biofilm formation supporting conditions for zoonotic serovars of Salmonella and avian pathogenic E. coli (APEC) from poultry have not been well studied yet. Clinical isolates of zoonotic Salmonella and APEC from poultry were evaluated for biofilm formation in four media at 37 degrees C and 40 degrees C after incubation of 48 and 72 hrs. The biofilms formed in 96 well plates were visualized and quantified with a new module of Aklides system using fluorescence microscope coupled with automated VideoScan Technology. After 72 hrs, brain heart infusion at 40 degrees C and Rappaport-Vassiliadis Soya broth at 37 degrees C were found most suitable for APEC and Salmonella biofilm formations respectively. The new information will be useful for further biofilm associated studies particularly for evaluation of antibiofilm compounds and contribute in infection control. (C) 2020 PVJ. All rights reserved
KW  - APEC
KW  - biofilm formation
KW  - Salmonella
KW  - video scan technology
Y1  - 2020
U6  - https://doi.org/10.29261/pakvetj/2020.066
SN  - 0253-8318
SN  - 2074-7764
VL  - 40
IS  - 4
SP  - 527
EP  - 530
PB  - University of Agriculture, Faculty of Veterinary Science
CY  - Faisalabad
ER  -