This study reports the rational design of broadly neutralizing human monoclonal antibodies (mAbs) against HIV-1. Researchers used knowledge of the HIV-1 envelope (Env) structure to create antigenically resurfaced glycoproteins that target the structurally conserved CD4-binding site (CD4bs) on gp120. These probes helped identify sera containing NAbs to the CD4bs and isolate B cells from an HIV-1-infected donor. By expressing immunoglobulin genes from individual cells, three mAbs were identified, including a pair of somatic variants that neutralized over 90% of circulating HIV-1 isolates. These mAbs, VRC01, VRC02, and VRC03, showed exceptional breadth of neutralization against diverse HIV-1 strains, including those from different genetic subtypes. The study demonstrates that targeting the functionally conserved CD4bs on gp120 can lead to broadly neutralizing antibodies, providing insights for future HIV-1 vaccine design. The research also highlights the importance of understanding the structural and functional aspects of the HIV-1 Env to develop effective vaccines. The findings suggest that a focused B cell response can target a highly conserved region of the HIV-1 Env, which is crucial for neutralization. The study also shows that these mAbs can be elicited in humans, offering new insights into how the human immune system can effectively target a vulnerable site on the viral Env. The results emphasize the potential of these mAbs for vaccine development and their ability to neutralize a wide range of HIV-1 strains.This study reports the rational design of broadly neutralizing human monoclonal antibodies (mAbs) against HIV-1. Researchers used knowledge of the HIV-1 envelope (Env) structure to create antigenically resurfaced glycoproteins that target the structurally conserved CD4-binding site (CD4bs) on gp120. These probes helped identify sera containing NAbs to the CD4bs and isolate B cells from an HIV-1-infected donor. By expressing immunoglobulin genes from individual cells, three mAbs were identified, including a pair of somatic variants that neutralized over 90% of circulating HIV-1 isolates. These mAbs, VRC01, VRC02, and VRC03, showed exceptional breadth of neutralization against diverse HIV-1 strains, including those from different genetic subtypes. The study demonstrates that targeting the functionally conserved CD4bs on gp120 can lead to broadly neutralizing antibodies, providing insights for future HIV-1 vaccine design. The research also highlights the importance of understanding the structural and functional aspects of the HIV-1 Env to develop effective vaccines. The findings suggest that a focused B cell response can target a highly conserved region of the HIV-1 Env, which is crucial for neutralization. The study also shows that these mAbs can be elicited in humans, offering new insights into how the human immune system can effectively target a vulnerable site on the viral Env. The results emphasize the potential of these mAbs for vaccine development and their ability to neutralize a wide range of HIV-1 strains.