In vitro immunization of mouse spleen cells for the production of monoclonal IgG1 antibodies using an antigen-specific T helper cell clone (D.10.G4.1)

Uthoff, Sonja M. S.; Böldicke, Thomas

doi:10.1016/0022-1759(93)90357-d

Cited by 12 publications

(3 citation statements)

References 36 publications

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“…Monoclonal mouse anti-ovalbumin and anti-avidin immunoglobulins were purchased from Sigma. Monoclonal mouse anti-ovotransferrin immunoglobulin was produced according to Uthoff and Bo ¨ldicke (16) and kindly supplied by Y. Nys and J. Gautron, INRA (Nouzilly, France). Secondary anti-mouse antibody was from Sigma.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Different Electrophoretic Separations of Hen Egg White Proteins

Désert¹,

Guérin-Dubiard²,

Nau³

et al. 2001

J. Agric. Food Chem.

104

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The hen egg white protein composition has not yet been fully defined. To improve the knowledge of this biological fluid, the most usual and recently developed electrophoretic methods have been used: SDS-PAGE, native-PAGE, isoelectric focusing (IEF), and 2-dimensional electrophoresis (2DE). Seven of the major known proteins were thus identified in at least one electrophoretic system. Isoforms of ovotransferrin, ovalbumin, and ovomucoid were visualized when pI was used for the separation. Two-dimensional electrophoresis allowed separation of a very large number of spots. In each of the four systems, some components were revealed but not identified, and unknown spots were particularly numerous with 2DE. With this technique, many spots corresponding to small acidic proteins were highlighted, among which was the Ch21 protein, whose presence in hen egg white was thus confirmed. This study thus constitutes, to our knowledge, the first proteomic investigation of hen egg white.

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Section: Introductionmentioning

confidence: 99%

Comparison of Different Electrophoretic Separations of Hen Egg White Proteins

Désert¹,

Guérin-Dubiard²,

Nau³

et al. 2001

J. Agric. Food Chem.

104

View full text Add to dashboard Cite

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“…In essence, the conditions in the germinal center (Victora and Nussenzweig, 2012) should be reproduced in vitro during immunization. The co-culture of splenocytes with follicular dendritic cells (Nishikawa et al, 2006) and T helper cell clones (Uthoff and Boldicke, 1993) are likely to activate B cells more efficiently. These stimulations would induce class switching and affinity maturation more effectively.…”

Section: Discussionmentioning

confidence: 99%

A method for inducing antigen-specific IgG production by in vitro immunization

Kato

Tsuji

et al. 2012

Journal of Immunological Methods

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“…Several research groups have developed various in vitro immunization procedures for the preparation of specific immortalized human B-lymphocytes using various fusion partners [ 14 , 15 ] or Epstein-Barr virus [ 16 – 18 ]. However, these techniques have a poor yield and reproducibility [ 19 – 21 ]. Furthermore, the protocols reported are complex since they require several steps for depletion or enrichment of various sub-populations [ 15 , 22 ].…”

Section: Introductionmentioning

confidence: 99%

Production of antigen-specific human IgGs by in vitro immunization

et al. 2016

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BackgroundWe previously developed in vitro immunization based on a fusion protein containing the transcriptional transactivator (Tat) of human immunodeficiency virus and a double domain, called ZZ, derived from protein A of Staphylococcus aureus. In this approach, naïve human peripheral blood mononuclear cells (PBMCs) trigger a specific IgM antibody (Ab) response in the presence of ZZTat. In the present study, we attempted to raise a specific IgG Ab response.ResultsWe found that PBMCs incubated with ZZTat and a mixture containing anti-CD40, IL4 and IL21 secrete anti-Tat IgG Abs in their supernatants, indicating that the cytokine cocktail provides an isotypic switch. Then, we deciphered the Tat determinant involved in the phenomenon and found that it is located in the region 22–57 and that, within this region, the cysteine-rich domain and the basic residues play a crucial role. Finally, we prepared a fusion protein containing a fragment derived from the NY-ESO-1 cancer/testis antigen (Ag) and showed that PBMCs incubated with ZZfNY-ESO-1Tat trigger a specific anti-fNY-ESO-1 IgG Ab response, which demonstrates the possibility of transferring immunizing ability to an Ag unrelated to Tat.ConclusionOur ZZTat-based in vitro immunization approach that offers the possibility to raise an IgG Ab response against NY-ESO-1 might represent a valuable first stage for the generation of fully human IgG specific Abs.

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In vitro immunization of mouse spleen cells for the production of monoclonal IgG1 antibodies using an antigen-specific T helper cell clone (D.10.G4.1)

Cited by 12 publications

References 36 publications

Comparison of Different Electrophoretic Separations of Hen Egg White Proteins

Comparison of Different Electrophoretic Separations of Hen Egg White Proteins

A method for inducing antigen-specific IgG production by in vitro immunization

Production of antigen-specific human IgGs by in vitro immunization

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