A butcher's son who deciphered the body's defense molecules
Alfred Nisonoff's story is not just one of scientific discovery, but of a unique mindset—a chemist's precision combined with an immunologist's vision. In the mid-20th century, when antibodies were mysterious molecules that somehow recognized and neutralized diseases, Nisonoff employed simple, elegant experiments to reveal their fundamental architecture. His work laid the structural groundwork for modern immunotherapy, making him a foundational figure in immunology whose insights continue to influence medicine today.
Born in 1923 in Corona, Queens, to Jewish immigrants from Hungary and Russia, Nisonoff grew up in South River, New Jersey, where his parents operated a butcher shop and grocery store1 .
Born in Corona, Queens to immigrant parents
Graduated high school at age 15
Graduated from Rutgers University
Served in U.S. Navy during WWII
Earned Ph.D. in biochemistry from Johns Hopkins
Passed away on March 12, 2001
In the late 1950s, key discoveries were emerging. Rodney Porter and Gerald Edelman were determining the fundamental structure of γ-globulins (now called immunoglobulins), discovering they consisted of two heavy and two light chains that could be cleaved into fragments4 .
Nisonoff's most famous experiment involved using pepsin to digest rabbit antibodies1 . While Porter's papain cleavage produced monovalent Fab fragments, Nisonoff discovered that pepsin cleaved antibodies differently—on the carboxyl-terminal side of the disulfide bonds connecting the two heavy chains1 .
Nisonoff's pepsin experiment didn't just reveal antibody structure—it opened the door to engineering antibodies with new functions. He discovered that the F(ab')₂ fragments could be broken down further into monovalent Fab' fragments through reduction, and these fragments could then be recombined under mild oxidation to form bivalent antibodies7 .
In a creative leap, Nisonoff mixed monovalent fragments from antibodies with two different specificities—anti-bovine gamma globulin (BGG) and anti-ovalbumin (OVA)—and allowed them to recombine2 . The result was the world's first bispecific antibody—a single molecule capable of binding two different antigens2 8 .
| Time Period | Key Development | Lead Scientists | Impact |
|---|---|---|---|
| Early 1960s | First bispecific F(ab')₂ fragments | Alfred Nisonoff | Proof of concept for bispecific antibodies2 8 |
| 1975 | Hybridoma technology | Köhler and Milstein | Enabled production of pure monoclonal antibodies2 8 |
| 1983 | Hybrid hybridoma (quadroma) | Milstein and Cuello | First method to produce full-length bispecific antibodies2 8 |
| 1996 | Knobs-into-holes technology | Genentech scientists | Solved heavy chain mispairing issue in production8 |
| 2009-2014 | First clinical approvals | Various companies | Catumaxomab (2009) and Blinatumomab (2014) approved for cancer8 |
Nisonoff's groundbreaking work relied on several key reagents and methods that became essential tools in immunology research.
| Reagent/Technique | Function in Research | Role in Nisonoff's Work |
|---|---|---|
| Papain | Protease enzyme that cleaves antibodies into Fab and Fc fragments | Used for comparative studies to understand pepsin's different cleavage site1 2 |
| Pepsin | Protease enzyme that cleaves antibodies into F(ab')₂ fragments | Key enzyme for producing bivalent antigen-binding fragments1 4 |
| Haptens | Small molecules that elicit antibody production when attached to carriers | Used to study antibody specificity and antigen binding1 7 |
| Reducing Agents | Chemicals that break disulfide bonds | Used to reduce F(ab')₂ to monovalent Fab' fragments7 |
| Oxidizing Conditions | Environment allowing reformation of disulfide bonds | Enabled recombination of Fab' fragments into bispecific antibodies2 7 |
| Quantitative Hapten Inhibition | Technique to measure antibody binding affinity | Extended Karl Landsteiner's work; used to study antibody specificity1 7 |
Nisonoff's career spanned decades and included leadership positions at the University of Illinois, Brandeis University, and numerous scientific organizations5 . He was elected to the National Academy of Sciences in 1984 and served as president of the American Association of Immunologists from 1990-19914 5 .
His 1975 monograph, "The Antibody Molecule," became the definitive reference work on the subject, synthesizing decades of antibody research into a comprehensive framework1 7 . Colleagues remembered him not just for his scientific rigor, but for his humility, collegiality, and straightforward approach to problem-solving4 .
Perhaps the most powerful testament to Nisonoff's work is its impact on modern medicine. The bispecific antibodies he first conceived in the early 1960s have become powerful cancer therapeutics. Blinatumomab, a bispecific T-cell engager approved by the FDA in 2014, has revolutionized treatment for certain types of acute lymphoblastic leukemia8 . By connecting a patient's T-cells to cancer cells, it effectively directs the immune system to destroy tumors—a concept Nisonoff first demonstrated in his laboratory decades earlier.
Alfred Nisonoff passed away on March 12, 2001, but his legacy endures in every bispecific antibody therapy, in our understanding of immunoglobulin structure, and in the continued exploration of the immune system's molecular machinery. His career exemplifies how simple, well-designed experiments can reveal profound biological truths—and how a chemist's perspective can forever change our understanding of life's processes.
"His goal was only to learn the truth." That he did, with both analytical brilliance and unexpected humility, leaving immunology forever in his debt4 .
Nisonoff authored numerous influential papers and the definitive monograph "The Antibody Molecule" in 1975.
Trained generations of immunologists and served in leadership roles at major institutions.
His discoveries laid the foundation for modern immunotherapy and bispecific antibody treatments.
Elected to the National Academy of Sciences and served as president of the American Association of Immunologists.