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Single domain antibodies: promising experimental and therapeutic tools in infection and immunity.

Wesolowski J, Alzogaray V, Reyelt J, Unger M, Juarez K, Urrutia M, Cauerhff A, Danquah W, Rissiek B, Scheuplein F, Schwarz N, Adriouch S, Boyer O, Seman M, Licea A, Serreze DV, Goldbaum FA, Haag F, Koch-Nolte F - Med. Microbiol. Immunol. (2009)

Bottom Line: VHH and VNAR are easily produced as recombinant proteins, designated single domain antibodies (sdAbs) or nanobodies.Other advantageous features of nanobodies include their small size, high solubility, thermal stability, refolding capacity, and good tissue penetration in vivo.Here we review the results of several recent proof-of-principle studies that open the exciting perspective of using sdAbs for modulating immune functions and for targeting toxins and microbes.

View Article: PubMed Central - PubMed

Affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

ABSTRACT
Antibodies are important tools for experimental research and medical applications. Most antibodies are composed of two heavy and two light chains. Both chains contribute to the antigen-binding site which is usually flat or concave. In addition to these conventional antibodies, llamas, other camelids, and sharks also produce antibodies composed only of heavy chains. The antigen-binding site of these unusual heavy chain antibodies (hcAbs) is formed only by a single domain, designated VHH in camelid hcAbs and VNAR in shark hcAbs. VHH and VNAR are easily produced as recombinant proteins, designated single domain antibodies (sdAbs) or nanobodies. The CDR3 region of these sdAbs possesses the extraordinary capacity to form long fingerlike extensions that can extend into cavities on antigens, e.g., the active site crevice of enzymes. Other advantageous features of nanobodies include their small size, high solubility, thermal stability, refolding capacity, and good tissue penetration in vivo. Here we review the results of several recent proof-of-principle studies that open the exciting perspective of using sdAbs for modulating immune functions and for targeting toxins and microbes.

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3D-structures of leukocyte ecto-enzymes and toxin enzymes with active site crevices targeted by sdAbs. a Murine T-cell ecto-ADP-ribosyltransferase ART2.2 with substrate NAD (pdb code 1og3). b Human lymphocyte ecto-NAD-glycohydrolase CD38 with substrate NAD (pdb code 2i65). cSalmonella enterica virulence protein SpvB, an actin ADP-ribosyltransferase, with NAD (pdb code 2gwl). dClostridium difficile toxin B, a rho glucosyltransferase (pdb code 2bvl). Images were generated with the PyMOL program [123]. We have already generated enzyme-blocking sdAbs against ART2.2 and SpvB from immunized llamas. Current work aims at generating similar antibodies against CD38 and Toxin B from immunized llamas and sharks
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Fig8: 3D-structures of leukocyte ecto-enzymes and toxin enzymes with active site crevices targeted by sdAbs. a Murine T-cell ecto-ADP-ribosyltransferase ART2.2 with substrate NAD (pdb code 1og3). b Human lymphocyte ecto-NAD-glycohydrolase CD38 with substrate NAD (pdb code 2i65). cSalmonella enterica virulence protein SpvB, an actin ADP-ribosyltransferase, with NAD (pdb code 2gwl). dClostridium difficile toxin B, a rho glucosyltransferase (pdb code 2bvl). Images were generated with the PyMOL program [123]. We have already generated enzyme-blocking sdAbs against ART2.2 and SpvB from immunized llamas. Current work aims at generating similar antibodies against CD38 and Toxin B from immunized llamas and sharks

Mentions: Small molecule inhibitors and antibodies offer two possible strategies for inhibiting leukocyte ecto-enzymes [1, 25]. While small chemical antagonists often lack specificity, antibodies have the capacity to discriminate between closely related enzymes. As leukocyte cell surface ecto-enzymes are accessible to antibodies, the remarkable preference of hcAbs for binding into the active sites of enzymes offers the possibility to raise selective enzyme-blocking sdAb-reagents [5, 6]. The crystal structures of several leukocyte ecto-enzymes have been elucidated, and in many cases they display a deep active site crevice that may be suitable for targeting by sdAbs (Fig. 8a, b).Fig. 8


Single domain antibodies: promising experimental and therapeutic tools in infection and immunity.

Wesolowski J, Alzogaray V, Reyelt J, Unger M, Juarez K, Urrutia M, Cauerhff A, Danquah W, Rissiek B, Scheuplein F, Schwarz N, Adriouch S, Boyer O, Seman M, Licea A, Serreze DV, Goldbaum FA, Haag F, Koch-Nolte F - Med. Microbiol. Immunol. (2009)

3D-structures of leukocyte ecto-enzymes and toxin enzymes with active site crevices targeted by sdAbs. a Murine T-cell ecto-ADP-ribosyltransferase ART2.2 with substrate NAD (pdb code 1og3). b Human lymphocyte ecto-NAD-glycohydrolase CD38 with substrate NAD (pdb code 2i65). cSalmonella enterica virulence protein SpvB, an actin ADP-ribosyltransferase, with NAD (pdb code 2gwl). dClostridium difficile toxin B, a rho glucosyltransferase (pdb code 2bvl). Images were generated with the PyMOL program [123]. We have already generated enzyme-blocking sdAbs against ART2.2 and SpvB from immunized llamas. Current work aims at generating similar antibodies against CD38 and Toxin B from immunized llamas and sharks
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2714450&req=5

Fig8: 3D-structures of leukocyte ecto-enzymes and toxin enzymes with active site crevices targeted by sdAbs. a Murine T-cell ecto-ADP-ribosyltransferase ART2.2 with substrate NAD (pdb code 1og3). b Human lymphocyte ecto-NAD-glycohydrolase CD38 with substrate NAD (pdb code 2i65). cSalmonella enterica virulence protein SpvB, an actin ADP-ribosyltransferase, with NAD (pdb code 2gwl). dClostridium difficile toxin B, a rho glucosyltransferase (pdb code 2bvl). Images were generated with the PyMOL program [123]. We have already generated enzyme-blocking sdAbs against ART2.2 and SpvB from immunized llamas. Current work aims at generating similar antibodies against CD38 and Toxin B from immunized llamas and sharks
Mentions: Small molecule inhibitors and antibodies offer two possible strategies for inhibiting leukocyte ecto-enzymes [1, 25]. While small chemical antagonists often lack specificity, antibodies have the capacity to discriminate between closely related enzymes. As leukocyte cell surface ecto-enzymes are accessible to antibodies, the remarkable preference of hcAbs for binding into the active sites of enzymes offers the possibility to raise selective enzyme-blocking sdAb-reagents [5, 6]. The crystal structures of several leukocyte ecto-enzymes have been elucidated, and in many cases they display a deep active site crevice that may be suitable for targeting by sdAbs (Fig. 8a, b).Fig. 8

Bottom Line: VHH and VNAR are easily produced as recombinant proteins, designated single domain antibodies (sdAbs) or nanobodies.Other advantageous features of nanobodies include their small size, high solubility, thermal stability, refolding capacity, and good tissue penetration in vivo.Here we review the results of several recent proof-of-principle studies that open the exciting perspective of using sdAbs for modulating immune functions and for targeting toxins and microbes.

View Article: PubMed Central - PubMed

Affiliation: Institute of Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

ABSTRACT
Antibodies are important tools for experimental research and medical applications. Most antibodies are composed of two heavy and two light chains. Both chains contribute to the antigen-binding site which is usually flat or concave. In addition to these conventional antibodies, llamas, other camelids, and sharks also produce antibodies composed only of heavy chains. The antigen-binding site of these unusual heavy chain antibodies (hcAbs) is formed only by a single domain, designated VHH in camelid hcAbs and VNAR in shark hcAbs. VHH and VNAR are easily produced as recombinant proteins, designated single domain antibodies (sdAbs) or nanobodies. The CDR3 region of these sdAbs possesses the extraordinary capacity to form long fingerlike extensions that can extend into cavities on antigens, e.g., the active site crevice of enzymes. Other advantageous features of nanobodies include their small size, high solubility, thermal stability, refolding capacity, and good tissue penetration in vivo. Here we review the results of several recent proof-of-principle studies that open the exciting perspective of using sdAbs for modulating immune functions and for targeting toxins and microbes.

Show MeSH
Related in: MedlinePlus