Computational Identification of Antigen-Binding Antibody Fragments

The calcium-binding protein S100A9 regulates inflammatory processes and the immune response. It is overexpressed in a variety of inflammatory and oncologic conditions. In this study, we produced a recombinant human S100A9 (hS100A9) antigen with high yield and purity and used it to generate a hybridoma cell culture-based monoclonal anti-hS100A9 antibody. We selected five anti-hS100A9 antibodies from cell supernatants that showed high antigen binding efficiency and identified the nucleotide sequences of three antibodies: two with high effective concentration values and one with the lowest value. The antigen and antibody development procedures described herein are useful for producing large amounts of monoclonal antibodies against hS100A9 and other antigens of interest. The nucleotide sequences of the anti-hS100A9 monoclonal antibody revealed herein will be helpful in the generation of recombinant antibodies or antibody fragments against hS100A9.

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The variability of nitro group-protein interaction in the 2,4-dinitrophenyl-binding antibodies M315, M460 and X25 investigated by resonance Raman spectroscopy

Biochemical Journal ◽

10.1042/bj1970119 ◽

1981 ◽

Vol 197 (1) ◽

pp. 119-125 ◽

Cited By ~ 2

Author(s):

P Gettins ◽

R A Dwek ◽

R N Perutz

Keyword(s):

Raman Spectroscopy ◽

Nitro Group ◽

Resonance Raman Spectroscopy ◽

Specific Interaction ◽

Resonance Raman ◽

Antibody Fragments ◽

Antibody Specificity ◽

Group Protein ◽

Binding Antibody ◽

Induced Changes

Pre-resonance Raman spectroscopy was used to study the interactions of the nitro groups of dinitrophenyl haptens with three dinitrophenyl-binding antibody fragments: M315 Fv, M460 Fab‘ and X25 Fab’. The observed changes in frequency of modes associated with the nitro moieties are compared with solvent-induced changes for the model hapten 2,4-dinitroaniline. These comparisons demonstrate a specific interaction via the H2N-C-C-2-NO2 and 4-NO2 groups with the protein. The interaction with the 4-NO2 group appears to be absent for epsilon-N-2,4-dinitrophenyl-L-lysine bound to M315 Fv fragment in contrast with either 2,4-dinitrophenylaspartate or 2,4-dinitrophenylglycine bound to M315 Fv fragment, despite the much tighter binding of the lysine derivative. The implications of this for M315 Fv fragment in terms of the antibody specificity are discussed. Comparisons of the effect of binding to M460 Fab‘ and X25 Fab’ fragments also revealed significant differences in the shifts of the nitro group vibrations of 2,4-dinitrophenyl-lysine and 2,4-dinitroaniline.

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Orientation of antigen binding sites in dimeric and trimeric single chain Fv antibody fragments

FEBS Letters ◽

10.1016/s0014-5793(98)00292-0 ◽

1998 ◽

Vol 425 (3) ◽

pp. 479-484 ◽

Cited By ~ 35

Author(s):

Lynne J. Lawrence ◽

Alexander A. Kortt ◽

Peter Iliades ◽

Peter A. Tulloch ◽

Peter J. Hudson

Keyword(s):

Binding Sites ◽

Antibody Fragments ◽

Antigen Binding ◽

Single Chain ◽

Single Chain Fv ◽

Fv Antibody ◽

Single Chain Fv Antibody

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Idiotypic self binding of a dominant germline idiotype (T15). Autobody activity is affected by antibody valency.

Journal of Experimental Medicine ◽

10.1084/jem.165.5.1332 ◽

1987 ◽

Vol 165 (5) ◽

pp. 1332-1343 ◽

Cited By ~ 25

Author(s):

C Y Kang ◽

H L Cheng ◽

S Rudikoff ◽

H Kohler

Keyword(s):

Sequence Analysis ◽

Binding Activity ◽

The Self ◽

Antigen Binding ◽

Simultaneous Presence ◽

Cdna Sequencing ◽

Antigen Binding Site ◽

Binding Antibody ◽

Polymeric State ◽

Germline Sequence

We have previously described (1-3) an IgM antibody that binds to PC, expresses the T15 idiotype, and binds also to itself or T15 if insolubilized. Because of the simultaneous presence of complementary idiotopes and paratopes this type of antibody has been termed autobody. The self binding involves the antigen-binding site because the F(ab')2 fragment of T15, PC, and no other haptens inhibit the self binding. DNA sequence analysis of 11E7-1 using primer extension cDNA sequencing showed that the variable sequences of H and L chains of 11E7-1 are identical to the germline sequence of the prototype T15 idiotype. Furthermore, monomeric and dimeric T15 IgA were shown to bind to insolubilized T15 and other T15+ antibodies including 11E7-1. Thus, the self-binding activity is an inherent property of the T15 germline sequence. The self binding is highly dependent on the polymeric state of the binding antibody since the IgM pentamer of 11E7-1 is about three fold more effective than the T15 dimer and 50 times more than the T15 monomer. These data suggest that the self-binding activity of a germline-encoded idiotype may play an important role in the biology of its expression, and more specifically, may be responsible for the establishment of its dominant expression.

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Transiently binding antibody fragments against Lewis x and sialyl-Lewis x

Journal of Immunological Methods ◽

10.1016/j.jim.2006.02.010 ◽

2006 ◽

Vol 312 (1-2) ◽

pp. 20-26 ◽

Cited By ~ 4

Author(s):

Reine Johansson ◽

Mats Ohlin ◽

Bo Jansson ◽

Sten Ohlson

Keyword(s):

Antibody Fragments ◽

Sialyl Lewis X ◽

Lewis X ◽

Sialyl Lewis ◽

Binding Antibody

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David vs. Goliath: The Structure, Function, and Clinical Prospects of Antibody Fragments

Antibodies ◽

10.3390/antib8020028 ◽

2019 ◽

Vol 8 (2) ◽

pp. 28 ◽

Cited By ~ 21

Author(s):

Adam Bates ◽

Christine A. Power

Keyword(s):

Monoclonal Antibodies ◽

Lower Cost ◽

Biological Effects ◽

Antibody Therapy ◽

Antibody Fragments ◽

Intracellular Targeting ◽

Fast Pace ◽

Binding Antibody ◽

Multifunctional Molecules ◽

Potential Use

Since the licensing of the first monoclonal antibody therapy in 1986, monoclonal antibodies have become the largest class of biopharmaceuticals with over 80 antibodies currently approved for a variety of disease indications. The development of smaller, antigen binding antibody fragments, derived from conventional antibodies or produced recombinantly, has been growing at a fast pace. Antibody fragments can be used on their own or linked to other molecules to generate numerous possibilities for bispecific, multi-specific, multimeric, or multifunctional molecules, and to achieve a variety of biological effects. They offer several advantages over full-length monoclonal antibodies, particularly a lower cost of goods, and because of their small size they can penetrate tissues, access challenging epitopes, and have potentially reduced immunogenicity. In this review, we will discuss the structure, production, and mechanism of action of EMA/FDA-approved fragments and of those in clinical and pre-clinical development. We will also discuss current topics of interest surrounding the potential use of antibody fragments for intracellular targeting and blood–brain barrier (BBB) penetration.

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