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Methods for producing human antibodies in SCID mice

a human antibody and mouse technology, applied in the field of human antibody production in scid mice, can solve the problems of complex previous methods, and achieve the effect of improving results and high serum antibody titers

Inactive Publication Date: 2006-07-20
COCCIA MARCO ANTHONY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The present invention is an improved method for producing human antibodies in mice. By using autologous dendritic cells and antigen-antibody complexes, the method results in high antibody titers with specificity. This method can also be used to produce human monoclonal antibodies to prostate specific antigen (PSA), which are useful for treating prostate cancer. These antibodies have human effector functions and are expected to elicit no immunogenicity."

Problems solved by technology

However, previous methods are typically very complex and yield inconsistent results.

Method used

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  • Methods for producing human antibodies in SCID mice
  • Methods for producing human antibodies in SCID mice
  • Methods for producing human antibodies in SCID mice

Examples

Experimental program
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Effect test

example 1

Human IgG Production in DC / Ab-PSA Complex Immunized

SCIDhu PBL Mice Mouse Monoclonal IgG.

[0038] Antibody (Cat. No. 10-P2O; Fitzgerald Industries Inc.) was complexed with PSA at equimolar ratios at 4° C. over night (Ab-PSA complex) and then dialyzed to remove azide. Autologous peripheral blood dendritic cells (pDC) were grown in serum free media and pulsed with either 25 μg / ml soluble PSA or Ab-PSA complex. All mice received 108 PBMC i.p. on day 0. Each group consisted of 8 mice. Group F mice were immunized with 25 g of Ab-PSA complex weekly and with 25 μg PSA on Day 21. Group G mice were immunized with 25 μg of soluble PSA weekly and 7×106 soluble PSA pulsed pDC on day 7. Group H mice were immunized with 25 μg of Ab-PSA complex weekly, 7.5×106 Ab-PSA pulsed pDC on day 7 and then 25 μg PSA on Day 21. These results are summarized in FIG. 6. The Day 14 results, (A) Graphs, from left to right are as follows: Average PSA specific IgG; PSA specific IgG for individual mice; Average tota...

example 2

Specificity of Antibody Responses Obtained in Ab-PSA / pDC Immunized SCIDhu PBC Mice.

[0043] In this Example, the relative PSA specificity of pooled sera from group H mice, control hIgG and group F serum is measured and was shown in FIG. 5.

[0044]FIG. 5 shows relative PSA specificity of pooled sera from group H mice, control hihg and group F serum. In this Figure, open circles represent pooled group H serum, and closed circles purified hIgG. The open triangles represent mouse F1.1 serum. The data shows that group H serum binds PSA 10 times greater than equal concentrations of either control hIgG. Panel B shows the inhibition of group H sera specific binding by soluble PSA. The open circles represent pooled group H serum diluted 1:15 (120 μg / ml). The closed circles represent pooled group H serum diluted 1;20 (90 μg / ml). The open tri-angles represent purified hIgG control, (90 μg / ml). The data shows that pooled group H sera binding can be inhibited by soluble PSA to OD490 values obtain...

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Abstract

An improved method for producing human antibodies in SCID mice is provided. The improvement includes the use of dendritic cells pulsed with antigen-antibody complexes and antigen-antibody complexes as immunizing agents.

Description

FIELD OF THE INVENTION [0001] The subject invention provides a novel and reproducible method for producing human monoclonal antibodies to desired antigens, e.g. prostate specific antigen. These monoclonal antibodies, because of their human origin, should be useful therapeutic agents, e.g. for the treatment of human prostate cancer. BACKGROUND OF THE INVENTION [0002] Antibodies (Ab) that recognize and adhere to proteins on the surface of bacteria, virus or parasites help immune system cells identify, attack and remove them from the body. Similarly, monoclonal Ab (MoAb) that adhere to cancer cells but not to normal cells can be an effective therapy for human cancers. Such MoAbs are generally murine Abs genetically modified to contain human constant regions (“humanized”). However, fully human MoAb are potentially superior to humanized murine MoAb as therapies for human cancer because of their absence of immunogenicity in humans. Human B cells can be stimulated to produce Abs that recog...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12P21/04A01K67/027A61K39/00C07K16/40
CPCA61K39/00A61K2039/5154A61K2039/5158C07K16/00C07K16/3069C07K16/40C07K2317/21A61K2239/38A61K2239/58A61K39/4622A61K39/464494A61K39/4615
Inventor COCCIA, MARCO ANTHONYBRAMS, PETER
Owner COCCIA MARCO ANTHONY