#antigen binding

Immunological Applications

Monoclonal antibodies bind:

Monoclonal antibody originates from unique genetic event where one developing B cell successfully completed VDJ recombination on heavy chain locus chromosome 14 mediated by RAG1/2 recognition of recombination signal sequences, non-homologous end joining with TdT adding N nucleotides creating unique complementarity determining region three loop that dominates antigen contact, plus VJ recombination on light chain locus producing paired variable domains. All progeny via mitosis carry identical nucleotide sequence encoding paratope whose three-dimensional shape, electrostatic surface, and hydropho

Ref: Abbas Cellular Molecular Immunology 10th ed monoclonal single epitope uniform affinity; Roitt Essential Immunology specificity.

View Details

Immunological Applications

Monoclonal antibodies bind:

Bevacizumab recombinant humanized IgG1 mAb 149 kDa neutralizes soluble VEGF-A isoforms 121 165 189 206 secreted tumor stromal inflammatory cells hypoxia HIF1alpha binding hypoxia response elements driving transcription. VEGF-A binds VEGFR1 FLT1 and R2 KDR tyrosine kinases endothelial cells dimerization autophosphorylation Tyr1175 recruiting PLC gamma generating IP3 DAG increasing calcium PKC stimulating proliferation permeability via VE-cadherin internalization gaps survival Akt phosphorylation caspase-9. Binding epitope Arg82 Lys84 affinity 0.5 nM overlapping receptor binding interface preven

Ref: Ferrara NEJM Bevacizumab VEGF Targeted Therapy 2004; FDA Avastin Mechanism Label VEGF Neutralization; Lodish Angiogenesis VEGF Signaling Pathway Chapter 23.

View Details

Microscopy

Which antibody fragment cannot precipitate?

Precipitation requires formation of large lattice through cross-linking where bivalent or multivalent antibodies bridge polyvalent antigens. Intact IgG with two Fab arms and F(ab')2 dimer retain bivalency, enabling network extension and insoluble aggregate generation. Whole antibody adds Fc but remains precipitable. In contrast, Fab fragment obtained by papain digestion is monovalent, possessing single binding site. It can bind epitope and block it, but cannot connect two antigen molecules to build lattice. Resulting complexes stay small and soluble, invisible in gel. Therefore Fab is used to

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

View Details

Microscopy

Which is true about Fab region?

Antibody molecule comprises Fab and Fc segments after papain cleavage. Fab includes entire light chain paired with variable and first constant domain of heavy chain. Hypervariable complementarity determining regions within variable domains fold to create paratope complementary to epitope shape, charge and hydrophobicity. This interaction determines specificity, affinity and neutralization capacity. Fc region mediates effector functions like complement fixation and Fc receptor engagement, not antigen recognition. All immunoglobulin isotypes and monoclonal preparations possess Fab, and it lacks

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

View Details