Abstract:
:A fragment of antigen binding (Fab) surface display system was developed using a glycoengineered Pichia pastoris host strain genetically modified to secrete glycoproteins with mammalian mannose-type Man(5)GlcNAc(2) N-linked glycans. The surface display method described here takes advantage of a pair of coiled-coil peptides as the linker while using the Saccharomyces cerevisiae Sed1p GPI-anchored cell surface protein as an anchoring domain. Several Fabs were successfully displayed on the cell surface using this system and the expression level of the displayed Fabs was correlated to that of secreted Fabs from the same glycoengineered host in the absence of the cell wall anchor. Strains displaying different model Fabs were mixed and, through cell sorting, the strain displaying more expressed Fab molecule or the strain displaying the Fab with higher affinity for an antigen was effectively enriched by FACS. This novel yeast surface display system provides a general platform for the display of Fab libraries for affinity and/or expression maturation using glycoengineered Pichia.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Lin S,Houston-Cummings NR,Prinz B,Moore R,Bobrowicz B,Davidson RC,Wildt S,Stadheim TA,Zha Ddoi
10.1016/j.jim.2011.10.003subject
Has Abstractpub_date
2012-01-31 00:00:00pages
159-65issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(11)00279-1journal_volume
375pub_type
杂志文章abstract::Household contacts of tuberculosis patients are at high risk of infection and development of active disease. In this study we evaluated the cytokine production and mRNA expression of IFN-γ, TNF-α, IL-10&IL-6 stimulated with r32kDa M. bovis BCGAg in active pulmonary tuberculosis patients (APTB), household contacts (HHC...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2016.02.012
更新日期:2016-05-01 00:00:00
abstract::A scheme for quantitation of serum and urinary paraproteins is described using isoelectric focusing and scanning densitometry. Paraproteins could be quantified using this system when present at concentrations ranging from 1 mg/ml to 26 mg/ml, depending on the immunoglobulin class. The relevance of these results to mon...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90082-7
更新日期:1986-06-24 00:00:00
abstract::An immunoperoxidase method has been developed which allows accurate and sensitive quantitation of the binding of monoclonal antibodies to cell surface antigens. Monolayers of fixed cells were prepared in wells of Terasaki micro-test plates and monoclonal antibodies bound to cell surface antigens were identified by the...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90240-9
更新日期:1980-01-01 00:00:00
abstract::Different numbers of Chinese hamster lymphocytes were cultured in microtiter plates with flat-, round- and V-bottomed wells for different culture times. The smallest number of cells could be stimulated in plates with V-bottomed wells. At least 25,000-50,000 cells and a longer culture time than for round- or V-bottomed...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90102-7
更新日期:1980-01-01 00:00:00
abstract::Stability of single-chain Fvs (scFvs) can be improved by mutagenesis followed by phage display selection where the unstable variants are first inactivated by, for example, denaturing treatment. Here we describe a modified strategy for the selection of stabilized antibody fragments by phage display, based on denaturati...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2004.11.008
更新日期:2005-01-01 00:00:00
abstract::The histamine content of commercial preparations of pokeweed mitogen was measured by amino acid analysis technique, automated fluorometry, and bioassay employing the guinea pig ileum. Ten samples from 5 companies were examined and found to contain between 0.026 micrograms and 167.5 micrograms of histamine per ml of so...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90095-3
更新日期:1982-12-30 00:00:00
abstract::Assessment of T cell activation has traditionally been performed by measuring proliferation as a function of 3[H]-thymidine incorporation, or secretion of cytokines from activated peripheral blood mononuclear cells (PBMC) in culture. An alternative method for detection of proliferation at the single cell level utilize...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)00127-0
更新日期:1997-10-13 00:00:00
abstract::A sensitive direct assay has been developed to quantitate antigen specific helper factors (ThF). The assay depends on the ability of ThF to block the binding of free antibody to immobilized antigen. It has been used to follow the course of purification of ThF produced in short-term cultures of in vitro primed helper c...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90098-x
更新日期:1984-02-24 00:00:00
abstract::The polymerase chain reaction (PCR) has proved to be a sensitive and versatile method for the analysis of human and murine cytokine mRNA expression. This paper describes for the first time a reverse transcription-polymerase chain reaction (RT-PCR) at end-point for the quantification of five porcine cytokines: interfer...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00105-2
更新日期:1999-10-29 00:00:00
abstract::Multiplex assays detecting sets of related clinical analytes simultaneously can save considerable amount of time and resources. Array-in-well (AIW) is a powerful platform for the multiplex detection of different analytes where microarrays can be printed at the bottom of microtiter wells, thus combining the potential o...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2015.12.007
更新日期:2016-02-01 00:00:00
abstract::A rapid and simple immunostick immunosorbent ELISA has been developed for the diagnosis of myasthenia gravis. It is based on coating immunosticks with the major myasthenia gravis autoantigens acetylcholine receptor (AChR) or MuSK and it can be performed in <1 h, in a non-specialized examination place by non-skilled pe...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2018.06.016
更新日期:2018-09-01 00:00:00
abstract::An automatic immunospecific affinity-chromatographic system for continuous operation is described. The system comprises time-controlled sample application, washing and elution steps and automatic dialysis of eluted fractions. The applicability of the system is illustrated by the purification of pregnancy zone protein ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(78)90117-5
更新日期:1978-01-01 00:00:00
abstract::The characterization of lymphoid subsets isolated from different anatomical sites is of great importance for understanding the mechanisms and interactions of normal and pathological immune reactions in the pig. The objective of this study was to standardize a protocol for the isolation of lymphocytes from mucosal tiss...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(00)00209-x
更新日期:2000-07-31 00:00:00
abstract::Two murine IgM monoclonal antibodies (MAs) directed against the lipid A portion of bacterial lipopolysaccharide (LPS) were compared in their binding to Re LPS and lipid A and their idiotypic make-up with rabbit anti-idiotypic sera. Horseradish peroxidase (HRPO)-labelled MAs 8-2 and 26-20 bound equally well to Re LPS. ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90308-i
更新日期:1990-06-12 00:00:00
abstract::A new method for measuring the uptake of Staphylococcus aureus by human polymorphonuclear neutrophils (PMN) using flow cytometry (FCM) is described. Bacteria were labeled with fluorescein isothiocyanate (FITC) and incubated with PMN in a suspension assay. At the end of the assay, phagocytosis was arrested by the addit...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(89)90161-0
更新日期:1989-07-26 00:00:00
abstract::Giant liposomes (mean diameter 5.5 microns) composed of egg phosphatidylcholine or distearoyl phosphatidylcholine, phosphatidyl glycerol, cholesterol and triolein were prepared by a double emulsion technique. They were then mixed with model particulate (killed Bacillus subtilis, and killed Bacille Calmette-Guérin) and...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90368-h
更新日期:1993-12-03 00:00:00
abstract::Two micro enzyme immunoassays (microEIA) for circulating immune complexes (CICs) are described. The Raji cell microEIA was similar in sensitivity, reproducibility and specificity to the Raji radioimmunoassay. The F(ab')2 anti-C3 microEIA was comparable to the 2 Raji cell assays. The 2 microEIAs for CICs were used to a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90258-3
更新日期:1985-11-07 00:00:00
abstract::Eosinophil cationic protein (ECP) is a highly basic and potent cytotoxic single-chain zinc-containing protein present in the granules of the eosinophilic granulocytes. ECP appears to be involved in defence against parasites and in the tissue damage seen in subjects with allergic and inflammatory disease. To investigat...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90177-h
更新日期:1991-04-25 00:00:00
abstract::Two sets of expression vectors were constructed that permitted the efficient expression of single-chain Fv fragments (scFvs) fused N-terminally to an enhanced mutant of the green fluorescent protein GFP+ or the red fluorescent protein DsRed in insect and mammalian cells. The vectors allowed rapid cloning of scFv fragm...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2003.12.001
更新日期:2004-02-15 00:00:00
abstract::We have developed and characterised a variant of the human T-cell tumour, MOLT-4F, which is resistant to both 8-azaguanine and 6-thioguanine. This new cell line, M4HS2, is suitable for the efficient production of lymphokine-secreting human T-cell hybrids and is available for distribution. ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90383-2
更新日期:1986-06-10 00:00:00
abstract::Exosomes are nanometer-sized vesicles, secreted from most cell types, with documented immune-modulatory functions. Exosomes can be purified from cultured cells but to do so effectively, requires maintenance of cells at high density in order to obtain sufficient accumulation of exosomes in the culture medium, prior to ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.03.001
更新日期:2008-06-01 00:00:00
abstract::Serum cytokine profiling is a powerful tool to link host immune defense with disease pathogenesis. Although several multiplex assays are commercially available, none has been rigorously validated in the context of chronic infectious disease (such as HIV infection). Here we compared the measurement of proinflammatory c...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2011.06.033
更新日期:2011-09-30 00:00:00
abstract::The study of T cell receptor (TCR) genes has been hampered by their large repertoires and elusive methods for gene amplification. We have developed a new method for amplification of all human TCR genes (alpha, beta, gamma, and delta) with the ligation of a universal adaptor to the leader sequence of variable (V) regio...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90193-b
更新日期:1993-05-05 00:00:00
abstract::Investigation of MHC class I endogenous antigen processing and presentation is important for understanding non-self immune recognition, vaccine design, immune evasion by pathogens and tumours, and elucidation of the mechanisms of certain auto-immune diseases. To study the MHC class I antigen-processing pathway, the pr...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.02.017
更新日期:2008-06-01 00:00:00
abstract::Flow cytometry offers a promising alternative to the current methods of amniocentesis or chorionic villus sampling (CVS) for fetal cell sorting for prenatal diagnosis. While flow cytometric methods have been greatly improved to be more sensitive at detecting fetal cells within the maternal circulation, there are still...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/j.jim.2010.09.039
更新日期:2011-01-05 00:00:00
abstract::A method is described for the simultaneous in situ detection of surface receptors and cytoplasmic granules in mast cells of frozen sections of mouse skin. Surface IgE receptors are detected after saturation of the receptors with a murine monoclonal antibody of IgE isotype. The latter is subsequently detected by monosp...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90121-9
更新日期:1987-05-20 00:00:00
abstract::The T cell receptor (TCR) determines the cellular response to antigens, which are presented on the surface of target cells in the form of a peptide bound to a product of the major histocompatibility complex (pepMHC). The response of the T cell depends on the affinity of the TCR for the pepMHC, yet many TCRs have been ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.09.016
更新日期:2008-12-31 00:00:00
abstract::The simian immunodeficiency virus (SIV) model of AIDS is widely used for the development of human immunodeficiency virus (HIV) vaccine strategies, particularly for the analysis of correlates of protective immunity. As it is not always possible to establish autologous B-lymphoblastoid cell lines (B-LCL) for use as targ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00480-x
更新日期:2001-12-01 00:00:00
abstract:OBJECTIVE:The objective of this study was the identification of the stain HIF-alpha using the Image Cytometry, and to help to count the positive cells (with HIF-alpha) and the negative cells (without HIF-alpha) from the same sample. METHOD:17 images of renal tissues from male rats of Winstar lineage; overall, there we...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2017.02.005
更新日期:2017-05-01 00:00:00
abstract::Typical clinical grade human IgG (intravenous immunoglobulin, IVIG), used for carbohydrate analysis, is derived from thousands of healthy donors. Quantitative high-resolution glycan profiles of IgG and its Fc-Fab fragments are presented here. Glycan profiles were established following digestions with Fc specific endog...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2012.05.022
更新日期:2012-08-31 00:00:00