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Immunology

Qualitative and Quantitative Assay for Detection of Circulating Autoantibodies against Human Aortic Antigen

Authors: Brent Veerman
Brent VeermanAffiliation: Department of Surgery, University of Toledo College of Medicine and Life Sciences, Toledo, USA
Bio-protocol author page: a4752
 and Ritu Chakravarti
Ritu ChakravartiAffiliation: Department of Surgery, University of Toledo College of Medicine and Life Sciences, Toledo, USA
For correspondence: ritu.chakravarti@utoledo.edu
Bio-protocol author page: a3737
,  date: 7/5/2017, 135 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2367.

[Abstract] Increased amount of autoantibodies in human sera are the hallmark of autoimmune diseases (Wang et al., 2015). In case of known antigen, detection of autoantibodies is done using laboratory based methods. However, in most autoimmune diseases, knowledge of self-antigen is still vague. We have developed an ELISA-based quantitative assay to detect the ...

Generation of IgG-Fc Glycovariants Using Recombinant Glycosidases and Glycosyltransferases

Authors: Isaak Quast
Isaak QuastAffiliation: Institute of Experimental Immunology, Department of Neuroinflammation, University Hospital of Zürich, Zürich, Switzerland
For correspondence: isaak.quast@uzh.ch
Bio-protocol author page: a3380
, Michael A. Maurer
Michael A. MaurerAffiliation 1: Institute of Experimental Immunology, Department of Neuroinflammation, University Hospital of Zürich, Zürich, Switzerland
Affiliation 2: Brain Research Institute, University of Zurich and Department of Health Sciences & Technology, ETH Zürich, Zürich, Switzerland
Bio-protocol author page: a3381
 and Jan D. Lünemann
Jan D. LünemannAffiliation: Institute of Experimental Immunology, Department of Neuroinflammation, University Hospital of Zürich, Zürich, Switzerland
Bio-protocol author page: a3382
,  date: 8/5/2016, 1578 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1886.

[Abstract] The immunoglobulin G (IgG) fragment crystallizable (Fc) domain contains a single, highly conserved asparagine 297 (N297) glycosylation site in the CH2 domain, which is buried within the hydrophobic core of each of the two heavy chains. The biantennary core glycan structure, composed of 2 N-acetylglucosamine (GlcNAc) and 3 mannose residues, can be further ...

Haemagglutination Inhibition (HI) Assay of Influenza Viruses with Monoclonal Antibodies

Authors: Ying Wu*
Ying WuAffiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a3175
, MyungSam Cho*
MyungSam ChoAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3176
, David Shore*
David ShoreAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3177
, Manki Song
Manki SongAffiliation: International Vaccine Institute, Seoul, Korea
Bio-protocol author page: a3178
, JungAh Choi
JungAh ChoiAffiliation: International Vaccine Institute, Seoul, Korea
Bio-protocol author page: a3179
, Tao Jiang
Tao JiangAffiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China
Bio-protocol author page: a3180
, Yong-Qiang Deng
Yong-Qiang DengAffiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China
Bio-protocol author page: a3181
, Melissa Bourgeois
Melissa BourgeoisAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3182
, Lynn Almli
Lynn AlmliAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3183
, Hua Yang
Hua YangAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3184
, Li-Mei Chen
Li-Mei ChenAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3185
, Yi Shi
Yi ShiAffiliation 1: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Affiliation 2: Research Network of Immunity and Health, Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a3186
, Jianxu Qi
Jianxu QiAffiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a3187
, An Li
An LiAffiliation 1: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Affiliation 2: College of Veterinary Medicine, Guangxi University, Nanning, China
Bio-protocol author page: a3188
, Kye Sook Yi
Kye Sook YiAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3189
, MinSeok Chang
MinSeok ChangAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3190
, Jin Soo Bae
Jin Soo BaeAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3191
, HyunJoo Lee
HyunJoo LeeAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3192
, JiYoung Shin
JiYoung ShinAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3194
, James Stevens
James StevensAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3195
, SeoungSuh Hong
SeoungSuh HongAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3196
, Cheng-Feng Qin
Cheng-Feng QinAffiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China
For correspondence: qincf@bmi.ac.cn
Bio-protocol author page: a3197
, George F. Gao
George F. GaoAffiliation 1: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Affiliation 2: Research Network of Immunity and Health, Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, China
Affiliation 3: Office of Director-General, Chinese Center for Disease Control and Prevention (China CDC), Beijing, China
For correspondence: gaof@im.ac.cn
Bio-protocol author page: a3198
, Shin Jae Chang
Shin Jae ChangAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
For correspondence: ShinJae.Chang@celltrion.com
Bio-protocol author page: a3199
 and Ruben O. Donis
Ruben O. DonisAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
For correspondence: rvd6@cdc.gov
Bio-protocol author page: a3200
 (*contributed equally to this work),  date: 6/5/2016, 2609 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1828.

[Abstract] Heamagglutination is inhibited when antibodies are present because antibodies to influenza virus will prevent attachment of the virus to red blood cells. The highest dilution of antibody that prevents hemagglutination is called the HI titer. The human monoclonal antibodies generated from single human B cells were tested to characterize their ability ...

Micro Neutralization (MN) Assay of Influenza Viruses with Monoclonal Antibodies

Authors: Ying Wu*
Ying WuAffiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a3175
, MyungSam Cho*
MyungSam ChoAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3176
, David Shore*
David ShoreAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3177
, Manki Song
Manki SongAffiliation: International Vaccine Institute, Seoul, Korea
Bio-protocol author page: a3178
, JungAh Choi
JungAh ChoiAffiliation: International Vaccine Institute, Seoul, Korea
Bio-protocol author page: a3179
, Tao Jiang
Tao JiangAffiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China
Bio-protocol author page: a3180
, Yong-Qiang Deng
Yong-Qiang DengAffiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China
Bio-protocol author page: a3181
, Melissa Bourgeois
Melissa BourgeoisAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3182
, Lynn Almli
Lynn AlmliAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3183
, Hua Yang
Hua YangAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3184
, Li-Mei Chen
Li-Mei ChenAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3185
, Yi Shi
Yi ShiAffiliation 1: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Affiliation 2: Research Network of Immunity and Health, Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a3186
, Jianxu Qi
Jianxu QiAffiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a3187
, An Li
An LiAffiliation 1: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Affiliation 2: College of Veterinary Medicine, Guangxi University, Nanning, China
Bio-protocol author page: a3188
, Kye Sook Yi
Kye Sook YiAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3189
, MinSeok Chang
MinSeok ChangAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3190
, Jin Soo Bae
Jin Soo BaeAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3191
, HyunJoo Lee
HyunJoo LeeAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3192
, JiYoung Shin
JiYoung ShinAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3194
, James Stevens
James StevensAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
Bio-protocol author page: a3195
, SeoungSuh Hong
SeoungSuh HongAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
Bio-protocol author page: a3196
, Cheng-Feng Qin
Cheng-Feng QinAffiliation: Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China
For correspondence: qincf@bmi.ac.cn
Bio-protocol author page: a3197
, George F. Gao
George F. GaoAffiliation 1: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
Affiliation 2: Research Network of Immunity and Health, Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, China
Affiliation 3: Office of Director-General, Chinese Center for Disease Control and Prevention (China CDC), Beijing, China
For correspondence: gaof@im.ac.cn
Bio-protocol author page: a3198
, Shin Jae Chang
Shin Jae ChangAffiliation: Biotechnology Research Institute, Celltrion, Inc., Incheon, South Korea
For correspondence: ShinJae.Chang@celltrion.com
Bio-protocol author page: a3199
 and Ruben O. Donis
Ruben O. DonisAffiliation: Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia, USA
For correspondence: rvd6@cdc.gov
Bio-protocol author page: a3200
 (*contributed equally to this work),  date: 6/5/2016, 1556 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1829.

[Abstract] The human monoclonal antibodies generated from single human B cells were tested to characterize their ability to neutralize virus infectivity. The microneutralization assay is a highly sensitive and specific assay for detecting virus-specific neutralizing antibodies to influenza viruses. This protocol is to measure the ability of human monoclonal antibody ...

Cell-based Assays to Monitor AID Activity

Authors: Ludivine C. Litzler
Ludivine C. LitzlerAffiliation 1: Institut de Recherches Cliniques de Montréal, Montréal, Canada
Affiliation 2: Department of Biochemistry, Université de Montréal, Montréal, Canada
Bio-protocol author page: a2889
, Stephen P. Methot
Stephen P. MethotAffiliation 1: Institut de Recherches Cliniques de Montréal, Montréal, Canada
Affiliation 2: Department of Medicine, McGill University, Montréal, Canada
Bio-protocol author page: a2890
, Anne-Marie Patenaude
Anne-Marie PatenaudeAffiliation: Institut de Recherches Cliniques de Montréal, Montréal, Canada
Bio-protocol author page: a2891
, Astrid Zahn
Astrid ZahnAffiliation: Institut de Recherches Cliniques de Montréal, Montréal, Canada
Bio-protocol author page: a2892
 and Javier M. Di Noia
Javier M. Di NoiaAffiliation 1: Institut de Recherches Cliniques de Montréal, Montréal, Canada
Affiliation 2: Department of Biochemistry, Université de Montréal, Montréal, Canada
Affiliation 3: Department of Medicine, McGill University, Montréal, Canada
For correspondence: Javier.Marcelo.Di.Noia@ircm.qc.ca
Bio-protocol author page: a2893
,  date: 2/5/2016, 1842 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1724.

[Abstract] The enzyme Activation induced deaminase (AID) underpins antibody affinity maturation and isotype switching through its mutagenic activity of deaminating deoxycytidine to deoxyuridine in DNA. Subsequent processing of the deoxyuridine initiates the processes of somatic hypermutation (SHM) and class switch recombination (CSR) in B cells. Structure-function ...

Coupling of HIV-1 gp120-derived Core Protein to Paramagnetic Beads and Adsorption Assays

Authors: Jidnyasa Ingale
Jidnyasa IngaleAffiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, USA
Bio-protocol author page: a2626
 and Richard T Wyatt
Richard T WyattAffiliation 1: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, USA
Affiliation 2: , IAVI Neutralizing Antibody Center at TSRI, La Jolla, USA
For correspondence: wyatt@scripps.edu
Bio-protocol author page: a2627
,  date: 10/5/2015, 1536 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1614.

[Abstract] Analysis of the functional activity in polyclonal serum following immunization of a complex protein or glycoprotein immunogen is a very important but tedious process. Fine mapping of epitope specific antibodies is difficult when they are elicited at relatively low levels. In our recent study focused at developing an HIV-1 vaccine, we immunized rabbits ...

Kinetic Analysis of Monoclonal Antibody Binding to HIV-1 gp120-derived Hyperglycosylated Cores

Authors: Jidnyasa Ingale
Jidnyasa IngaleAffiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, USA
Bio-protocol author page: a2626
 and Richard T Wyatt
Richard T WyattAffiliation 1: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, USA
Affiliation 2: , IAVI Neutralizing Antibody Center at TSRI, La Jolla, USA
For correspondence: wyatt@scripps.edu
Bio-protocol author page: a2627
,  date: 10/5/2015, 1602 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1615.

[Abstract] Kinetic analysis of antibodies is one of the important study for characterization of antibodies and screening of ligands. In our recent study1, we compared the antigenic profiles and binding characteristics of four HIV-1 envelope glycoprotein (Env) core immunogens using multiple monoclonal antibodies by Bio-Layer Light Interferometry (BLI). This technology ...

Fluorescence-linked Antigen Quantification (FLAQ) Assay for Fast Quantification of HIV-1 p24Gag

Authors: Marianne Gesner
Marianne GesnerAffiliation: Gladstone Institutes, University of California (UCSF), San Francisco, USA
Bio-protocol author page: a1891
, Mekhala Maiti
Mekhala MaitiAffiliation: Gladstone Institutes, University of California (UCSF), San Francisco, USA
Bio-protocol author page: a1892
, Robert Grant
Robert GrantAffiliation: Gladstone Institutes, University of California (UCSF), San Francisco, USA
Bio-protocol author page: a1893
 and Marielle Cavrois
Marielle CavroisAffiliation: Gladstone Institutes, University of California (UCSF), San Francisco, USA
For correspondence: mcavrois@gladstone.ucsf.edu
Bio-protocol author page: a1583
,  date: 12/20/2014, 2567 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1366.

[Abstract] The fluorescence-linked antigen quantification (FLAQ) assay allows a fast quantification of HIV-1 p24Gag antigen. Viral supernatant are lysed and incubated with polystyrene microspheres coated with polyclonal antibodies against HIV-1 p24Gag and detector antibodies conjugated to fluorochromes (Figure 1). After washes, the fluorescence of microspheres ...

Binding Affinity Measurement of Antibodies from Crude Hybridoma Samples by SPR

Authors: Dorin Mlaki Ndao
Dorin Mlaki NdaoAffiliation: AC Immune SA, Lausanne, Switzerland
Bio-protocol author page: a1751
, David T. Hickman
David T. HickmanAffiliation: AC Immune SA, Lausanne, Switzerland
Bio-protocol author page: a1752
, María Pilar López-Deber
María Pilar López-DeberAffiliation: AC Immune SA, Lausanne, Switzerland
Bio-protocol author page: a1753
, Aurélien Davranche
Aurélien DavrancheAffiliation: AC Immune SA, Lausanne, Switzerland
Bio-protocol author page: a1754
, Andrea Pfeifer
Andrea PfeiferAffiliation: AC Immune SA, Lausanne, Switzerland
Bio-protocol author page: a1755
 and Andreas Muhs
Andreas MuhsAffiliation: AC Immune SA, Lausanne, Switzerland
For correspondence: andreas.muhs@acimmune.com
Bio-protocol author page: a1756
,  date: 11/5/2014, 4561 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1276.

[Abstract] Surface Plasmon Resonance (SPR) is widely used to generate kinetic and affinity information on specific interactions between biomolecules. This technique is label-free and monitors the binding event in real-time. It is generally used for characterization of monoclonal antibody - antigen interactions. This protocol describes specifically the use of ...

Pulse Chase of Suspension Cells

Authors: Lai-Yee Wong
Lai-Yee WongAffiliation: Department of Molecular Microbiology and Immunology, University of Southern California, Los Angeles, CA, USA
For correspondence: laiyee.wong@usc.edu
Bio-protocol author page: a1453
, QiMing Liang
QiMing LiangAffiliation: Department of Molecular Microbiology and Immunology, University of Southern California, Los Angeles, CA, USA
Bio-protocol author page: a1454
, Kevin Brulois
Kevin BruloisAffiliation: Department of Molecular Microbiology and Immunology, University of Southern California, Los Angeles, CA, USA
Bio-protocol author page: a1455
 and Jae Jung
Jae JungAffiliation: Department of Molecular Microbiology and Immunology, University of Southern California, Los Angeles, CA, USA
Bio-protocol author page: a1456
,  date: 7/5/2014, 3746 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1170.

[Abstract] Pulse-chase method is a powerful technique used to follow the dynamics of proteins over a period of time. The expression level, processing, transport, secretion or half-life of proteins can be tracked by metabolically labeling the cells, such as with radiolabeled amino acids (pulse step). This protocol describes the condition used to study the folding ...
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INF-gamma Release ELISpot Assay

Author: Huagang Zhang
Huagang ZhangAffiliation: Albert Einstein College of Medicine, Yeshiva University, New York City, USA
For correspondence: huagangzhang@gmail.com
Bio-protocol author page: a21
,  date: 3/20/2012, 9504 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.120.

[Abstract] IFN-γ (Interferon-gamma) is produced mainly by activated T cells and NK cells. Production of IFN-γ (Interferon-gamma) by helper T cells as well as cytotoxic T cells is a hallmark of the TH1-type phenotype, thus, high-level production of IFN-γ (Interferon-gamma) is typically associated with effective ...

Antibody Purification from Western Blotting

Author: Lin Fang
Lin FangAffiliation: Department of Pediatrics, School of Medicine, Stanford University, Stanford, USA
For correspondence: cheerfulfang@hotmail.com
Bio-protocol author page: a20
,  date: 3/20/2012, 9417 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.133.

[Abstract] This protocol describes a method of purifying antibodies from sera with denatured antigens immobilized on western blot membranes. Advantages include (1) fast and easy; (2) purification of antibody with antigen in denatured form allows high yield in case antigen protein solubility is limited. Disadvantage ...

[Bio101] Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Murine Anti-dsDNA Antibodies

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
,  Anne Davidson Lab, date: 2/20/2011, 8297 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.32.

[Abstract] ELISA is an easy and relatively sensitive way to measure protein concentration. This protocol describes how to measure serum levels of murine anti-dsDNA antibodies. Circulating anti-dsDNA antibody is a hallmark of SLE both in human patients and in many SLE mouse models. This protocol was developed or ...

[Bio101] Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Total Murine Immunoglobulin

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
,  Anne Davidson Lab, date: 2/5/2011, 8234 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.33.

[Abstract] ELISA is an easy and relatively sensitive way to measure protein concentration. This protocol describes how to measure serum levels of murine total immunoglobulin of various isotypes. This protocol was developed or modified in Dr. Anne Davidson’s lab at Feinstein Institute for Medical Research. ...

ELISpots Assay to Measure the Number of Murine Plasma Cells Producing Anti-dsDNA Antibodies

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
,  date: 1/5/2012, 7556 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.34.

[Abstract] Circulating anti-dsDNA antibody is a hallmark of SLE both in human patients and in many SLE mouse models. This protocol describes how to measure the frequency of plasma cells producing these antibodies in the spleen of lupus-prone mice. This protocol was developed or modified in Dr. Anne Davidson’s ...

Isolation of Circulating Immune Complexes from TB Patient Serum for Serodiagnosis

Authors: Uma Devi Ranganathan
Uma Devi RanganathanAffiliation: Department of Pediatrics (Infectious Diseases), Albert Einstein College of Medicine, Bronx, USA
For correspondence: krumadevi@gmail.com
Bio-protocol author page: a44
, Ramalingam Bethunaickan
Ramalingam BethunaickanAffiliation: Department of Autoimmune and Musculoskeletal Diseases, Feinstein Institute for Medical Research, Manhasset, USA
Bio-protocol author page: a24
 and Alamelu Raja
Alamelu RajaAffiliation: Department of Immunology, Tuberculosis Research Center (ICMR), Chennai, India
Bio-protocol author page: a223
,  date: 6/5/2012, 6657 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.188.

[Abstract] Estimation of circulating immune complex in tuberculosis patients has shown better insight to the infection. Isolating circulating immune complex helps quantifying both antigen and antibody in the serum. It’s a simple procedure to improve the sensitivity and specificity in serodiagnosis of tuberculosis. ...

Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Murine Anti-cardiolipin Antibodies

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
,  date: 6/20/2012, 6449 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.227.

[Abstract] The circulating anticardiolipin antibody is a hallmark of antiphospholipid syndrome. It also appears in a number of autoimmune mouse models and is indicative of the break of tolerance against self antigens. This protocol describes a reliable method to determine the relative serum titer of anticardiolipin ...

[Bio101] Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Murine Anti-histone Antibodies

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
,  date: 6/20/2012, 5921 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.228.

[Abstract] Autoimmune disease is characterized with the break of tolerance against self antigens. Anti-histone antibodies can be found in the majority of patients with system lupus erythematosus (SLE) and a number of lupus-prone mouse strains. This protocol describes a reliable method to determine the relative ...

[Bio101] ELISA Measurement of Mouse IL-2

Author: Guo N. Huang
Guo N. HuangAffiliation: Program in Biochemistry, Cellular and Molecular Biology, Johns Hopkins University School of Medicine, Baltimore, USA
For correspondence: guo.n.huang@gmail.com
Bio-protocol author page: a30
,  date: 1/5/2012, 5595 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.169.

[Abstract] Interleukin-2 (IL-2) is a cytokine secreted by T cells that is essential for immune system activation. This protocol is routinely used for quantification of IL-2 concentration in the supernant of cultured lymphocytes under various stimulations and co-culturing conditions. Upon slightly modification ...

Monoclonal Antibody Purification (Nicotiana benthamiana Plants)

Authors: Adam Husk
Adam HuskAffiliation: Owensboro Cancer Research Program at James Graham Brown Cancer Center, University of Louisville School of Medicine, Owensboro, USA
Bio-protocol author page: a1126
, Krystal Teasley Hamorsky
Krystal Teasley Hamorsky Affiliation: Owensboro Cancer Research Program at James Graham Brown Cancer Center, University of Louisville School of Medicine, Owensboro, USA
Bio-protocol author page: a1127
 and Nobuyuki Matoba
Nobuyuki MatobaAffiliation: Department of Pharmacology and Toxicology, and Owensboro Cancer Research Program at James Graham Brown Cancer Center, University of Louisville School of Medicine, Owensboro, USA
For correspondence: n.matoba@louisville.edu
Bio-protocol author page: a1128
,  date: 1/20/2014, 4998 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1034.

[Abstract] Plant-based expression systems provide an alternative biomanufacturing platform for recombinant proteins (Matoba et al., 2011). In particular, plant virus-based vectors can overexpress proteins within days in the leaf tissue of Nicotiana benthamiana (N. benthamiana). To overcome the issues of genetic ...
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